Human Papillomavirus E6E7 mRNA and TERC lncRNA in Situ Detection in Cervical Scraped Cells and Cervical Disease Progression Assessment

Human Papillomavirus E6E7 mRNA and TERC lncRNA in Situ Detection in Cervical Scraped Cells and Cervical Disease Progression Assessment | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Human Papillomavirus E6E7 mRNA and TERC lncRNA in Situ Detection in Cervical Scraped Cells and Cervical Disease Progression Assessment Hui Zhao, Yue He, Bei Fan, Yan Wang, Yu-Mei Wu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-838435/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 24 Jan, 2022 Read the published version in Virology Journal → Version 1 posted 13 You are reading this latest preprint version Abstract Background Human papillomavirus screen in female cervical cells has demonstrated values in clinical diagnosis of precancerous lesions and cervical cancers. Human papillomavirus tests of cervical cells by utilizing Polymerase Chain Reaction (PCR) method provides human papillomavirus infection status however no further virus in situ information. Although it is well known that the tests of human papillomavirus E6/E7 RNA location in infected cervical cells and cell internal malignancy molecular will provide clues for gynecologists to evaluate disease progression, there are technique difficulties to preserve RNAs in cervical scraped cells for in situ hybridization. Methods In current study, after developing a cervical cell collection and preparation methods for RNA in situ hybridization, we captured the chance to screen 98 patient cervical cell samples and detected human papillomavirus E6/E7 mRNAs of high-risk subtypes, low-risk subtypes and lncRNA TERC in the cells. Results There are 70% consistence between human papillomavirus PCR and human papillomavirus RNA in situ hybridization results in cervical collected cells. Viral E6/E7 mRNAs were observed to distribute in cervical cell nuclear and cytoplasm. Moreover, viral gathered clusters were observed outside of cells through human papillomavirus RNA in situ hybridization. Varied numbers of human papillomavirus infective cells were detected by RNAscope assay in different patients even though they are all human papillomavirus high-risk subtype positive discovered by human papillomavirus PCR results. A cell malignancy related long non-coding RNA, TERC, has been detected in seven patient samples. The patient follow-up information was further analyzed with RNAscope results which indicated a combination of RNAscope positive signals of TERC and human papillomavirus high risk signals in more than 10 cells (cytoplasm or nucleus) may connect with cervical lesion fast progression which deserves further studies in the future. Conclusions Taken together, current study has provided an observable clue for gynecologists to evaluate human papillomavirus infection stage and cell malignancy status which may contribute for assessment of cervical disease progression. Virology RNA in situ hybridization HPV E6/E7 cervical scraped cells cervical malignancy tests Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Human papillomavirus (HPV) is a group of more than 200 related viruses, which are widely spread through vaginal, anal or oral sex. It has been proven that HPV can cause multiple types of cancers including cervical cancers and head and neck cancers. More than 9 of every 10 cases of cervical cancer are caused by HPV which brings the fact that monitor HPV status benefiting the diagnosis of precancers and cervical cancers. Given the fact that cervical cancer is once the leading causes of cancer deaths among women worldwide, the HPV detection becomes a key test for cervical diseases. Among eight genes HPVs encode, only E6 and E7 have transforming properties [1]. E6 and E7 oncoproteins are necessary for malignant conversion by associating with tumor suppressors p53 and pRB, respectively, to promote cell proliferation. Methods trying to detect HPV in cervical area cells have been widely established to monitor cervical disease. Since no robust IHC assays for HPV E6 and E7 are available, polymerase chain reaction (PCR) methods targeting on HPV whole genome have been routinely used for HPV subtype identification. Although DNA based PCR are sensitive to detect HPV subtypes, it is impossible to visualize HPV transcripts in cells and tissues which is further disabled to understand active HPV viral infection amount, viral subcellular locations and cervical cell transforming status. In order to include cell context information into HPV tests, p16 IHC has been developed as a surrogate marker based on the findings that HPV E7 oncoprotein binds to Rb protein region which leads to p16 overexpression [2]. I n situ detection of HPV E6 and E7 mRNA becomes available after RNAscope technology has been developed. As a novel generation of RNA in situ hybridization, this method is designed to detect E6/E7 RNAs of different HPV subtypes, for example, HPV HR-18 has included 18 high risk of HPV subtypes (HPVs 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82). The test utilizes 10 pairs of oligonucleotide probes per HPV type with amplification after probe hybridization [3]. With the conquering of HPV oncogene E6 and E7 in situ detection method by RNAscope technology, it is available to study active HPV infective status in cervical samples. In the past several years, studies have focused on HPV E6/E7 mRNA features on cervical pathological samples to support clinical diagnosis [1] , however there is no active HPV viral in situ tests performed in patient cervical scraped cells due to failed RNA preservation in such samples. In order to follow up HPV viral status and its relation of cervical lesion in cervical scraped cells, we have developed a new protocol to enable cervical scraped cells fitting for RNAscope HPV study. HPV E6/E7 RNA in situ information has been investigated in 98 patients’ cervical scraped cell samples to understand HPV E6/E7 RNA distribution in patient cervix, its correlation with HPV qPCR results, and patient disease progression. Our study discovered 70% consistence between RNAscope and PCR results. HPV E6/E7 RNA signals showed varied distribution pattern either in cytoplasm, in cell nucleus or as clusters gathered outside of cervical cells. RNAscope results of TERC, a long non-coding RNA (lncRNA), were co-tested to further evaluate cell malignance in the same HPV tested samples. Materials And Methods Ethics approval and consent to participate This study was conducted with the approval of the Beijing Obstetrics and Gynecology hospital, Capital Medical University, Institutional Review Board (IRB) Committee on Human Research in the Medical Sciences (CHRMS). A written informed patient consent was signed by each patient before joining this study project. All agrees to provide specimens and their data to be further published as part of the study results. Patient population and sample preparations 115 adult female outpatients ranging in age from 23 to 71 years were included in this study. The cervical samples were collected between Dec 2018 and March 2019 followed the procedures described below. After exposure cervical entrance, its surface was scratched two circles to collect cervical scraped cells using two different TCT sample collection brushes, respectively. For most patients, first circle of scraped cells was sent for HPV PCR tests, whereas the second circle of scraped cells was tested for RNAscope assays. For the latter collected ones, the TCT sample collection brush was cut and the tip with cells were kept in a 50ml tube with 10% neutral buffered formalin (NBF). The tubes were kept at 4 º C overnight then the cell samples on the brush tips were physically scraped down from the brush into 10% NBF. The tubes were centrifuged at 800rmp for 10min to collect cells. Cell pellet of each sample were transferred into 2ml EP tube and washed by PBS once. 1.5ml 70% ethanol was used to resuspend the cells and stored at room temperature (RT) for 2hr. The cells in 2ml EP tubes were then centrifuged at 8000-10000 rpm for 5min and the supernatant was discarded. The cell pellets were regarded as a chunk and went through 70%, 80%, 95% and 100% ethanol, respectively, at RT for 10min. After 100% ethanol, the cell pellet chunk floated and were transferred into a filter paper to totally try. Melt CellGel (Beijing Pursuit Bio Co., ltd.) was dropped onto hydrophobic paper (parafilm paper). The dried cell pellet chunk was embedded in the CellGel, solidified with the CellGel and became a bigger block. The latter one was transferred into tissue processing histology Cassette and went through 85% ethanol for 45 min; 95% ethanol for 30min, 100% ethanol I for 30min, 100% ethanol II for 30min, 100% ethanol III for 45 min, xylene I for 30min, xylene II for 30min, xylene III for 45min, Wax I for 30min, Wax II for 30min, Wax III for 30min and Wax IV for 30min, then embedded in paraffin to become Formalin-fixed paraffin-embedding (FFPE) blocks. Each FFPE block carried a patient cervical cell pellet were sectioned of 5μm for RNA in situ hybridization tests. RNA Chromogenic in situ Hybridization RNA in situ hybridization was performed on FFPE cell pellet sections (5 μm) using the RNAscope 2.5 HD assay-Red (Advanced Cell Diagnostics, Inc.) and the RNAscope Probes (Advanced Cell Diagnostics, Inc.) including HPV-HR18 (pool of 18 individual high-risk human papillomavirus subtype E6/E7 mRNA probes: HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, and 82), HPV-LR6 (pool of 6 individual low-risk HPV subtype E6/E7 mRNA probes 6, 11, 40, 42, 43 and 44) and Hs-TERC probe. A negative probe targeting diaminopimelate B (DapB) and a positive RNA probe targeting human ubiquitin C (Hs-UBC), were used to evaluate each sample quality. Samples with no signal from DapB, and score >= 2 by UBC were counted as quality control (QC) passed. The RNAscope 2.5 HD-Red manual assays was followed per the manufacturer’s instructions. Each sample was tested for RNA quality control (QC) firstly (Hs-UBC and DapB). The QC passed ones were further studied using HPV-HR18, HPV-LR6 and Hs-TERC probes, respectively. qPCR Analysis of HPV DNA Human Papillomavirus Polymerase Chain Reaction HR-HPV PCR was performed using the 23 HPV Genotyping Real-time PCR Kit (Hybribio, China) containing 17 high risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73 and 82, and 6 low risk HPV types: 6, 11, 42, 43, 44 and 81. Interpreting Results RNAscope stained FFPE cell sections were scanned using Leica AT2 scanner (Leica, US). Whole sections were examined at 40× magnification. RNAscope results of HPV were recorded based on signal location and positive cells. For probe-HPV-HR and probe-HPV-LR results, the signal locations of cells and the positive cell numbers in each sample, 10, were recorded. Besides classic RNAscope signals in cytoplasm or nuclear, there are HPV RNAs gathered as clusters above one or more cells, which were recorded as well. For TERC results, RNA signals in nucleus were recorded. Two gynecologists (Z. H. and H. Y.) evaluated the scanned sections independently. If a disagreement occurred, they reviewed the case together and reached the final agreement. The interpretation was generally straightforward; therefore, no significant disagreements lead to incompatibility. Results In current study, totally 115 patient cervical cell samples have been collected. 101 samples have been successfully prepared into FFPE blocks and passed RNAscope positive control tests using the probe of Hs-UBC. Three of 101 samples failed RNAscope negative control tests with background of probe DapB staining. All 98 QC passed samples were studied using RNAscope HPV high-risk probe (V-HPV-HR18) and HPV low-risk probe (V-HPV-LR6). Hs-TERC probe targeting on cell malignance has also been detected in the samples. Among 98 analyzed patient samples, 69 showed consistent results between RNAscope and PCR from patient cervical scraped cells. 46 of 69 patient samples were PCR and RNAscope both negative (supplemental table 1). 23 of 69 patient samples were PCR and RNAscope both positive of HPV high risk subtypes (table 1). Among them, 15 patients showed positive HPV RNAscope signals in cells (cytoplasm or nucleus) with or without RNA signal clusters located outside (above) of cells (figure 1, a-c), whereas another 8 samples only carried HPV RNA signals as clusters located out of cells (table 1) (figure 1, d-f). Five patient samples exhibited lncRNA TERC signals in cell nucleus which indicated cell transformation signs (figure 2). The rest 29 patient samples with inconsistent RNAscope and PCR results were further divided into 3 subgroups (table 2). The first subgroup included 8 patient samples which were HPV RNAscope positive, either high risk or low risk subtypes, however PCR HPV negative. In this subgroup, RNAscope results indicated 4 samples carried HPV RNA signals in cells (figure 3, a), whereas the other 4 carried HPV RNA signals as clusters above cells (table 2-1) (figure 3, b-e). Notably four samples in this subgroup came from patients with medical history of post-surgery of Loop Eelectrosurgical Excision Procedure (LEEP) or at Cervical Intraepithelial Neoplasia (CIN) I/II level, respectively. Moreover, all RNAscope positive signals limited in small number of cells (≦10) no matter signal exhibition phenotype (table 2-1). The 2 nd subgroup included 9 samples which were PCR results positive however RNAscope negative (table 2-2). One patient sample exhibited TERC RNAscope signals with a history of CIN II and post-surgery of Leep for 3 years (figure 3, f). The last subgroup consisted of 12 patients which were HPV positive detected by both RNAscope and PCR methods, however inconsistent between high-risk and low-risk subtypes (table 2-3) (figure 4, a-e). Four patients had medical history of CKC treatment, laser CO2 vaporization therapy or CINIII, respectively. One patient (patient #48) in the subgroup had no previous HPV infection history, her sample exhibited HPV high-risk RNAscope signals in cytoplasm (>10) and HPV low risk RNAscope clusters. PCR results showed HPV 52 positive. Moreover, the patient TERC RNAscope signals were positive (figure 4, f). Discussion Previous studies have confirmed that p16 expression is associated with cervical lesion classification [4], namely the heavier the degree of cervical lesions is, the higher the degree of p16 expresses. As a parameter to judge the disease severity, p16 indicates surgery necessity when the lesion has been developed to certain stages. Gynecologists would prefer a parameter which could predict the progression of cervical lesion, especially when the lesion is at the initial stage or even no lesions are there yet. In China, immunohistochemistry of p16 and ki67 are mostly performed on pathological samples which need invasive performance, e.g. colposcopy, accompanied with bleeding and infections sometimes. For women during pregnancy, cervical colposcopy biopsy caused bleeding may stimulate contractions. It may increase the risk of miscarriage or premature delivery and increase the psychological burden during the gestation period. For no pregnancy patients, if the lesions are located at the cervical canal, commonly happened in postmenopausal women, it is hard to reach by colposcopy. In order to obtain the biopsy of those areas, cone cutting surgeries are commonly selected which will increase patient unnecessary injury burden, with the risk of missed diagnosis still. It is necessary to find a method to assess the disease severity without colposcopy and even predict the risk of cervical disease progression. The ideal assay results should be able to determine whether a colposcopy biopsy is unavoidable for patient with high-risk subtype of HPV infection. High-risk subtypes of HPV infection are known to have chance to lead cervical cells into transforming stages and may have more chance to develop into high squamous intraepithelial lesion (HSIL) [5]. HPV PCR tests are therefore widely performed in clinical to monitor cervical lesion. With the fact that HPV PCR results examine HPV DNAs without the information of viral activity status, there are HPV PCR positive samples carried no active virus from patients who may be overjudged for aggressive treatments or misestimated disease procession. Moreover, HPV infective cell numbers and the location information of HPV active virus in/near cervical cells are lacking by PCR tests. It is reasonable to estimate different disease progression if active HPV high risk viral RNAs have been detected widely spread in huge number of cells vs. only in several cells. Current study is the first report trying to estimate cervical lesion progression by using RNAscope HPV in situ hybridization in cervical scraped cells as we have known. By developing a new method which solved the main technique difficulties of cell detachment and RNA quality issue of cervical scraped cells, it makes RNA in situ hybridization of HPV E6/E7 become feasible. The results uncovered a window to study active HPV infection status and how the viral locally interacts with cervical cells which provides observable clues for disease progression assessment. 98 patient samples which passed RNAscope QC have been examined by using HPV high-risk probes, RNAscope HPV low-risk probes and TERC probe which is a long non-coding RNA associated with high grade squamous intraepithelial neoplasia (HSIL) and progression of invasive carcinoma. RNAscope results have been compared to PCR assay data for further analysis. In most cases, cervical scraped cells were collected twice, the former set was used for PCR assays and the latter set was used for RNAscope assays. In several cases, PCR used cell samples were collected at different date based on patients’ situation. The collection time variations may partly lead to the 29 inconsistence of HPV results between RNAscope and PCR assays. HPV DNA results detected by PCR study were compared to active HPV transcribed E6/E7 mRNAs captured by RNAscope assays, which may also lead to varied results if HPV virus infection happened previously however with inactive status. RNAscope assay result demonstrated two types of HPV signals. One is typical RNA signal dots located in cell nuclear and cytoplasm. The other type of signals exhibited big clusters located above cells, which looks like “out of cell” signals. The latter signal phenotype has been observed in both high-risk and low-risk HPV probe detected results which demonstrate viral secreting status. In the patients whose sample were RNAscope positive whereas PCR negative, 4 were after-LEEP or CIN I-III stage. RNAscope positive signals in the samples were most out of cells, with only 4 patient samples showed HPV positive signals in nucleus (<10). In nine RNAscope negative whereas PCR positive samples, seven were HPV High-risk subtypes. RNAscope results negative may due to sample and tested cell variations, inactive virus status or other sample preparation caused unknown reasons. Five of ninety-eight samples (patient 1, 5, 20, 26 and 48) exhibited active HPV E6/E7 mRNA signals in more than 10 cells (supplemental table 2). Moreover, patient 20 and 48 were TERC positive. Four (patient 1, 20, 26, 48) were high-risk HPV subtypes. Follow up records indicate that patient 1, 20 and 48 have received Leep (patient 1 and 20) and hysterectomy (patient 48), respectively, shortly after this study. To be noticed, patient 48 was detected HPV 52 positive by PCR tests with no infection history, whereas RNAscope discovered HPV high-risk subtype positive with more than 10 infective cells, plus TERC positive. Except Patent 20 and 48, there are five more patient (patient 40, 60, 61, 75 and 77) were TERC positive by RNA in situ tests. Among them, patient 40, 60 and 77 carried HPV high- risk E6/E7 mRNA more than 3 cervical cells (supplemental table 2). Follow up records indicated that patient 40 and 77 had lost contact since 2019. Patient 60 follow-up records showed HPV low risk positive only (2020 May). Patient 61 follow-up records showed HPV PCR 52 positive (2020 June). To be noticed, patient 61 had received LEEP (2016. Oct) before the study starting. By 2019 Dec, patient 75 was both HPV PCR and TCT negative (supplemental table 2). Conclusion Current results suggest that a combination of RNAscope positive signals of TERC and HPV high-risk subtype signals in more than 10 cells (cytoplasm or nucleus) may connect with cervical lesion fast progression which deserves highly attention. Abbreviations Human papillomavirus (HPV) Polymerase Chain Reaction (PCR) long non-coding RNA (lncRNA) Institutional Review Board (IRB) Committee on Human Research in the Medical Sciences (CHRMS) neutral buffered formalin (NBF) room temperature (RT) Formalin-fixed paraffin-embedding (FFPE) diaminopimelate B (DapB) human ubiquitin C (Hs-UBC) quality control (QC) Loop Eelectrosurgical Excision Procedure (LEEP) Cervical Intraepithelial Neoplasia (CIN) high squamous intraepithelial lesion (HSIL) Declarations Acknowledgement We thank the scientists of Advanced Cell Diagnostics, Inc., Dr. Na Li and her team (Mr. Zhifu Zhang, Mrs. Ying Zhou, Mrs. Hongzhe Sun and Mrs. Xin Wang) for technical assistance. Author’s contributions Dr. Yu-Mei Wu designed the study and data analysis. Dr. Zhao Hui wrote the manuscript, performed patient sample collections, did RNA in situ experiments and data analysis. Dr. Yue He, Bei Fan and Yan Wang all participated patient sample collections and RNA in situ tests. Dr. Yue He did RNA in situ data scoring. Funding This study was supported by Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support (grant no. ZYLX201705). Availability of data and material All detailed data and methods and material have been included in the material and method sections and result sections. Ethics approval and consent to participate This study was conducted with the approval of the Beijing Obstetrics and Gynecology hospital, Capital Medical University, Institutional Review Board (IRB) Committee on Human Research in the Medical Sciences (CHRMS). Consent for publication Informed consent was obtained from all individual participants included in the study. Conflict of interest statement This article has no conflict of interest References 1. Wardak S: Human Papillomavirus (HPV) and cervical cancer Med Dosw Mikrobiol 2016, 68 73-84. 2. Major AL, Dvořák V, Schwarzová J, Skřivánek A, Malík T, Pluta M, Mayboroda I, Grandjean EM: Efficacy and safety of an adsorbent and anti-oxidative vaginal gel on CIN1 and 2, on high-risk HPV, and on p16/Ki-67: a randomized controlled trial. Arch Gynecol Obstet 2020 Nov 20, doi: 10.1007/s00404-020-05816-8 . 3. Evans MF, Peng Z, Clark KM, Adamson CS-C, Ma X-J, Wu X, Wang H, Luo Y, Cooper K: HPV E6/E7 RNA in situ hybridization signal patterns as biomarkers of three-tier cervical intraepithelial neoplasia grade. PLoS One 2014, 9: e91142. 4. Sano T, Oyama T, Kashiwabara K, Fukuda T, Nakajima T: Expression Status of p16 Protein Is Associated with Human Papillomavirus Oncogenic Potential in Cervical and Genital Lesions. The American Journal of Pathology 1998, 153,: 1741-1748. 5. Regauer S, Reich O, Kashofer K: Thin variant of high-grade squamous intraepithelial lesion - relationship with high-risk and possibly carcinogenic human papilloma virus subtypes and somatic cancer gene mutations. Histopathology 2019 75: 405-412. Tables Table 1 Comparison of RNAscope and PCR results in patient cervical scraped cells. The detection results are consistent between the two methods. RNAscope results PCR results patient # Probe V-HPV-HR18 Note Probe V-HPV-LR6 Note Probe Hs-TERC Note HPV subtype Note Age sample collector P#1 P cyto and nucleus, >10 N N 16, 53, 56, 58 ASCUS 39 Doctor #1 P#8 P cyto, 10 N P 16 CINII-III 42 Doctor #1 P#22 P cyto and nucleus, >=3 N N 52 47 Doctor #1 P#24 P cyto and nucleus, >=3 N N 58 postsurgery of CKC 30 Doctor #2 P#28 P cyto and nucleus, >=3 N N 51 LSIL 32 Doctor #1 P#39 P cyto, =3 N P Not test Early invasion in cervical carcinoma 63 Doctor #1 P#41 P cyto and nucleus, >=3 N N 52 postsurgery of LEEP 47 Doctor #3 P#47 P cyto, >=3 N N 18 28 Doctor #1 P#60 P cyto, >=3 N P 53 HPV infection history 25 Doctor #2 P#69 P cyto and nucleus, >=3 N N 52 23 Doctor #4 P#77 P nuclear, >=3 N P 82, 42 42 Doctor #1 P#82 P cyto and nucleus, >=3 N N 6, 59 58 Doctor #2 P#83 P cyto and nucleus, >=3 N N 53 CINI 65 Doctor #4 P#49 P cluster, >=3 N N 51 postsurgery of LEEP 2 years 32 Doctor #1 P#57 P cluster, <3 N N 53 HPV infection history and CINI 29 Doctor #2 P#71 P cluster, <3 N N 52 CINI 45 Doctor #4 P#75 P cluster, <3 N P 52 CINII 50 Doctor #4 P#81 P cluster, <3 N N 33 50 Doctor #1 P#89 P cluster, <3 Y cluster, =3 N N 56 61 Doctor #2 P#71 P cluster, <3 N N 52 CINI 45 Doctor #4 Tale 2-1 Comparison of RNAscope and PCR results in patient cervical scraped cells- RNAscope positive whereas PCR negative. RNAscope results PCR results patient # Probe V-HPV-HR18 Description Probe V-HPV-LR6 Description Probe Hs-TERC Description HPV subtype Note age sample collector P#2 P cluster, 3 N N postsurgery of Loop Eelectrosurgical Excision Procedure (LEEP) 34 Doctor #1 P#18 N P cluster, =3 N N N postsurgery of Loop Eelectrosurgical Excision Procedure (LEEP) 45 Doctor #2 P#23 N P nucleus, >=3 N N 49 Doctor #2 P#32 P nucleus, <3 P cluster, =3 P cluster, >=3 N N CINII，postsurgery of Loop Eelectrosurgical Excision Procedure(LEEP) 2 years 32 Doctor #4 P#67 P nucleus and cluster, >=3 N N N CINI Cervical Intraepithelial Neoplasia 38 Doctor #4 P#80 P cluster, <3 N N N 28 Doctor #1 Table 2-2 Comparison of RNAscope and PCR results in patient cervical scraped cells- RNAscope negative whereas PCR positive. RNAscope results PCR results patient # Probe V-HPV-HR18 Description Probe V-HPV-LR6 Description Probe Hs-TERC Description HPV subtype Note age sample collector P#10 N N N 51, 52 34 Doctor #1 P#15 N N N 39 36 Doctor #1 P#17 N N N 58 CKC after 27 Doctor #1 P#27 N N N 33, 52, 58 64 Doctor #3 P#54 N N N 33 32 Doctor #4 P#61 N N P nucleus, >10 52 CINI-II, postsurgery of Leep (3 years) 60 Doctor #4 P#62 N N N 81 CINI 53 Doctor #4 P#100 N N N 44 45 Doctor #4 P#96 N N N 16 VaINIII, Vaginal Intraepithelial Neoplasia 71 Doctor #1 Table 2-3 Comparison of RNAscope and PCR results in patient cervical scraped cells- RNAscope and PCR both positive whereas inconsistence between high-risk and low-risk subtypes. RNAscope results PCR results patient # Probe V-HPV-HR18 Description Probe V-HPV-LR6 Description Probe Hs-TERC Description HPV subtype Note age sample collector P#4 P nuclear, >=3 P nuclear, >=3 N 53, 56 postsurgery of Cold Knife Conization(CKC) 53 Doctor #1 P#5 N P cyto, >10 N 11, 59 62 Doctor #1 P#6 N P cluster, <3 N 68, 43 postsurgery of Cold Knife Conization(CKC) 27 Doctor #1 P#9 P cluster, =3 N 43 69 Doctor #1 P#19 N P cluster, 10 N N 44, 45 CINIII 39 Doctor #1 P#30 N P cluster, 10 N 56 56 Doctor #1 P#33 P cluster, <3 P cluster, 10 P cluster, >10 P nucleus, >10 52 60 Doctor #1 P#53 N P cyto, <3 N 45 35 Doctor #4 P#73 P cluster, <3 N N 42, 43 42 Doctor #1 Supplementary Files HISPatientConsentForm.pdf InsitudetectionofHPVE6E7supplementalinformation.docx Cite Share Download PDF Status: Published Journal Publication published 24 Jan, 2022 Read the published version in Virology Journal → Version 1 posted Editorial decision: Major revision 03 Oct, 2021 Review # 3 received at journal 02 Oct, 2021 Review # 2 received at journal 01 Oct, 2021 Review # 1 received at journal 26 Sep, 2021 Reviewer # 3 agreed at journal 18 Sep, 2021 Reviewer # 2 agreed at journal 17 Sep, 2021 Reviews received at journal 17 Sep, 2021 Reviewers invited by journal 17 Sep, 2021 Reviewer # 1 agreed at journal 16 Sep, 2021 Submission checks completed at journal 06 Sep, 2021 Editor invited by journal 03 Sep, 2021 Editor assigned by journal 03 Sep, 2021 First submitted to journal 22 Aug, 2021 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-838435","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":51475347,"identity":"546c82aa-eda4-4020-aea4-aa8ef597db1d","order_by":0,"name":"Hui Zhao","email":"","orcid":"https://orcid.org/0000-0003-2093-6458","institution":"Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Beijing Maternal and Child Health Care Hospital","correspondingAuthor":false,"prefix":"","firstName":"Hui","middleName":"","lastName":"Zhao","suffix":""},{"id":51475348,"identity":"e85caca9-e735-43fc-be2f-a57d7a78dc61","order_by":1,"name":"Yue He","email":"","orcid":"","institution":"Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Beijing Maternal and Child Health Care Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yue","middleName":"","lastName":"He","suffix":""},{"id":51475349,"identity":"ac7aa8ed-675e-4c7e-b3dc-36668fc51e53","order_by":2,"name":"Bei Fan","email":"","orcid":"","institution":"Beijing Obstetrics and Gynecology Hospital, Capital Medical Univerrsity. Beijing Maternal and Child Healthy Care Hospital","correspondingAuthor":false,"prefix":"","firstName":"Bei","middleName":"","lastName":"Fan","suffix":""},{"id":51475350,"identity":"150b6690-de37-4fdd-a56d-1f165933e519","order_by":3,"name":"Yan Wang","email":"","orcid":"","institution":"Beijing Obstetriics and Gynecology Hospital, Capital Medical University. Beijing Maternal and Child Healthy Care Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yan","middleName":"","lastName":"Wang","suffix":""},{"id":51475351,"identity":"0e42513a-22f9-4dd3-9384-2e6eb6320749","order_by":4,"name":"Yu-Mei Wu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6klEQVRIiWNgGAWjYHAC9g8f/0nw8LM3QPkHCGthY5zBZiEj2QNTSowWZh62ChuDGwlEajG4kZ32mIdHgkdy5uOHn262Mcjx3Uhg/FyAV0vudsM5EkC/SKcZS+e2MRhL3khglp6BR4vZjdwNEm8MgLbMzmFjBmpJ3HAjAehUQlp4EiR4DG6eAWupJ0bLNkmeA0AtN3jAWhIMCGmxP/N2s+HMBqDDeoB+yTknYTjzzMNmaXxaJNtzNz742FBnz89++OHnnDIbeb7jyQc/49PCIJCAwpUAYsYGfBoYGPgP4JcfBaNgFIyCUcAAAJunSuxEACIPAAAAAElFTkSuQmCC","orcid":"","institution":"Beijing Obstetriics and Gynecology Hospital, Capital Medical University. Beijing Maternal and Child Healthy Care Hospital","correspondingAuthor":true,"prefix":"","firstName":"Yu-Mei","middleName":"","lastName":"Wu","suffix":""}],"badges":[],"createdAt":"2021-08-23 02:27:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-838435/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-838435/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12985-021-01696-9","type":"published","date":"2022-01-24T10:51:46+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":13320174,"identity":"ea213ef7-fa2c-428e-ab77-1b2eed5979fb","added_by":"auto","created_at":"2021-09-13 15:24:21","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":16875387,"visible":true,"origin":"","legend":"RNAscope HPV-HR18 representative images. a-f, Patient #20 (a), #40 (b), #28(c), #75 (d), #49 (e) and #57 (f). HPV PCR results of the patients were positive. ","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/bf2dde16c35bed5d57bdfaf3.jpg"},{"id":13320171,"identity":"7628fa4c-2513-4b5e-8348-c4831e02abc0","added_by":"auto","created_at":"2021-09-13 15:24:20","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":13784323,"visible":true,"origin":"","legend":"RNAscope Hs-TERC representative images. a-e, Patient #77 (a), #20 (b), #60 (c), # 75 (d), #40 (e). HPV PCR results of the patients were all HPV-high risk positive. ","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/9b2c1e837e728b6625d0d140.jpg"},{"id":13320173,"identity":"e6c91463-8d15-4931-bc30-dc1f8921bcfc","added_by":"auto","created_at":"2021-09-13 15:24:20","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":17362572,"visible":true,"origin":"","legend":"RNAscope HPV-HR18, HPV-LR6 and Hs-TERC representative images. a-f, RNAscope HPV-HR18 representative images of Patient #21 (a), #2 (b), #67 (c) and #59 (d). HPV-LR6 representative image of patient #2 (e) and Hs-TERC representative image of patient #61 (f). HPV PCR results of the patients were all negative. ","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/936a2c87dd90685917163cfb.jpg"},{"id":13320172,"identity":"df897f9b-4e88-4c20-a92a-1afbbe444989","added_by":"auto","created_at":"2021-09-13 15:24:20","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":16733067,"visible":true,"origin":"","legend":"RNAscope HPV-HR18, HPV-LR6 and Hs-TERC representative images. a-f, Patient #4 (a), #4 (b), #6 (c), #48 (d), #26 (e) and #48 (f), HPV PCR results of the patients were positive, whereas with different HPV subtypes from RNAscope results. ","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/c885dc4b2a45c704030ea917.jpg"},{"id":17584982,"identity":"f8af0495-f465-42a5-9bf6-5655a1e6fcae","added_by":"auto","created_at":"2022-01-24 10:51:49","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":991960,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/19b3b201-609c-434c-990b-e60d87034496.pdf"},{"id":13320170,"identity":"3496f01b-5fa7-4f14-9634-eb0723eed917","added_by":"auto","created_at":"2021-09-13 15:24:20","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":70208,"visible":true,"origin":"","legend":"","description":"","filename":"HISPatientConsentForm.pdf","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/6beba6c05fb74a339dfb0264.pdf"},{"id":13320169,"identity":"8a5121ea-b628-43c1-9dc5-804eedc26514","added_by":"auto","created_at":"2021-09-13 15:24:20","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":30782,"visible":true,"origin":"","legend":"","description":"","filename":"InsitudetectionofHPVE6E7supplementalinformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-838435/v1/297614717f91673b2b26a1d4.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eHuman Papillomavirus E6E7 mRNA and TERC lncRNA in Situ Detection in Cervical Scraped Cells and Cervical Disease Progression Assessment\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eHuman papillomavirus (HPV) is a group of more than 200 related viruses, which are widely spread through vaginal, anal or oral sex. It has been proven that HPV can cause multiple types of cancers including cervical cancers and head and neck cancers. More than 9 of every 10 cases of cervical cancer are caused by HPV which brings the fact that monitor HPV status benefiting the diagnosis of precancers and cervical cancers. Given the fact that cervical cancer is once the leading causes of cancer deaths among women worldwide, the HPV detection becomes a key test for cervical diseases.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong eight genes HPVs encode, only E6 and E7 have transforming properties\u0026nbsp;[1]. E6 and E7 oncoproteins are necessary for malignant conversion by associating with tumor suppressors p53 and pRB, respectively, to promote cell proliferation. Methods trying to detect HPV in cervical area cells have been widely established to monitor cervical disease. Since no robust IHC assays for HPV E6 and E7 are available, polymerase chain reaction (PCR) methods targeting on HPV whole genome have been routinely used for HPV subtype identification. Although DNA based PCR are sensitive to detect HPV subtypes, it is impossible to visualize HPV transcripts in cells and tissues which is further disabled to understand active HPV viral infection amount, viral subcellular locations and cervical cell transforming status. In order to include cell context information into HPV tests, p16 IHC has been developed as a surrogate marker based on the findings that HPV E7 oncoprotein binds to Rb protein region which leads to p16 overexpression\u0026nbsp;[2]. \u003cem\u003eI\u003c/em\u003e\u003cem\u003en situ\u0026nbsp;\u003c/em\u003edetection of HPV E6 and E7 mRNA becomes available after RNAscope technology has been developed. As a novel generation of RNA\u003cem\u003e\u0026nbsp;in situ\u0026nbsp;\u003c/em\u003ehybridization, this method is\u0026nbsp;designed to detect\u0026nbsp;E6/E7\u0026nbsp;RNAs of different HPV subtypes, for example, HPV HR-18 has included 18 high risk of HPV subtypes (HPVs 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82). The test utilizes 10 pairs of oligonucleotide probes per HPV type with amplification after probe hybridization\u0026nbsp;[3].\u0026nbsp;With the conquering of HPV oncogene E6 and E7 \u003cem\u003ein situ\u003c/em\u003e detection method by RNAscope technology, it is available to study active HPV infective status in cervical samples.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn the past several years, studies have focused on HPV E6/E7 mRNA features on cervical pathological samples to support clinical diagnosis\u0026nbsp;[1]\u0026nbsp;, however there is no active HPV viral \u003cem\u003ein situ\u0026nbsp;\u003c/em\u003etests performed in patient cervical scraped cells due to failed RNA preservation in such samples. In order to follow up HPV viral status and its relation of cervical lesion in cervical scraped cells, we have developed a new protocol to enable cervical scraped cells fitting for RNAscope HPV study. HPV E6/E7 RNA \u003cem\u003ein situ\u003c/em\u003e information has been investigated in 98 patients\u0026rsquo; cervical scraped cell samples to understand HPV E6/E7 RNA distribution in patient cervix, its correlation with HPV qPCR results, and patient disease progression. Our study discovered 70% consistence between RNAscope and PCR results. HPV E6/E7 RNA signals showed varied distribution pattern either in cytoplasm, in cell nucleus or as clusters gathered outside of cervical cells. RNAscope results of TERC, a long non-coding RNA (lncRNA), were co-tested to further evaluate cell malignance in the same HPV tested samples. \u0026nbsp;\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cem\u003eEthics approval and\u0026nbsp;\u003c/em\u003e\u003cem\u003econsent to participate\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThis study was conducted with the approval of the\u0026nbsp;Beijing Obstetrics and Gynecology hospital, Capital Medical University,\u0026nbsp;Institutional Review Board (IRB) Committee on Human Research in the Medical Sciences (CHRMS). A written informed patient consent was signed by each patient before joining this study project.\u0026nbsp;All agrees to provide specimens and their data to be further published as part of the\u0026nbsp;study results.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003ePatient population and sample preparations\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e115 adult female outpatients ranging in age from 23 to 71 years were included in this study. The cervical samples were collected between Dec 2018 and March 2019 followed the procedures described below. After exposure cervical entrance, its surface was scratched two circles to collect cervical scraped cells using two different TCT sample collection brushes, respectively. For most patients, first circle of scraped cells was sent for HPV PCR tests, whereas the second circle of scraped cells was tested for RNAscope assays. \u0026nbsp;For the latter collected ones, the TCT sample collection brush was cut and the tip with cells were kept in a 50ml tube with 10% neutral buffered formalin (NBF). The tubes were kept at 4\u003csup\u003e\u0026ordm;\u003c/sup\u003eC overnight then the cell samples on the brush tips were physically scraped down from the brush into 10% NBF. The tubes were centrifuged at 800rmp for 10min to collect cells. Cell pellet of each sample were transferred into 2ml EP tube and washed by PBS once. 1.5ml 70% ethanol was used to resuspend the cells and stored at room temperature (RT) for 2hr. The cells in 2ml EP tubes were then centrifuged at 8000-10000 rpm for 5min and the supernatant was discarded. The cell pellets were regarded as a chunk and went through 70%, 80%, 95% and 100% ethanol, respectively, at RT for 10min. After 100% ethanol, the cell pellet chunk floated and were transferred into a filter paper to totally try. Melt CellGel\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e(Beijing Pursuit Bio Co., ltd.) was dropped onto hydrophobic paper (parafilm paper). The dried cell pellet chunk was embedded in the CellGel, solidified with the CellGel and became a bigger block. The latter one was transferred into tissue processing histology Cassette and went through 85% ethanol for 45 min; 95% ethanol for 30min, \u0026nbsp;100% ethanol I for 30min, 100% ethanol II for 30min, 100% ethanol III for 45 min, xylene I for 30min, xylene II for 30min, xylene III for 45min, Wax I for 30min, Wax II for 30min, Wax III for 30min and Wax IV for 30min, then embedded in paraffin to become Formalin-fixed paraffin-embedding (FFPE) blocks. Each FFPE block carried a patient cervical cell pellet were sectioned of 5\u0026mu;m for RNA \u003cem\u003ein situ\u003c/em\u003e hybridization tests.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eRNA Chromogenic in situ Hybridization\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eRNA \u003cem\u003ein situ\u003c/em\u003e hybridization was performed on FFPE cell pellet sections (5 \u0026mu;m) using the RNAscope 2.5 HD assay-Red (Advanced Cell Diagnostics, Inc.) and the RNAscope Probes (Advanced Cell Diagnostics, Inc.) including HPV-HR18 (pool of 18 individual high-risk human papillomavirus subtype E6/E7 mRNA probes: HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, and 82), HPV-LR6 (pool of 6 individual low-risk HPV subtype E6/E7 mRNA probes 6, 11, 40, 42, 43 and 44) and Hs-TERC probe. A negative probe targeting diaminopimelate B (DapB) and a positive RNA probe targeting human ubiquitin C (Hs-UBC), were used to evaluate each sample quality. Samples with no signal from DapB, and score \u0026gt;= 2 by UBC were counted as quality control (QC) passed. The RNAscope 2.5 HD-Red manual assays was followed per the manufacturer\u0026rsquo;s instructions. Each sample was tested for RNA quality control (QC) firstly (Hs-UBC and DapB). The QC passed ones were further studied using HPV-HR18, HPV-LR6 and Hs-TERC probes, respectively.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eqPCR Analysis of HPV DNA\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHuman Papillomavirus Polymerase Chain Reaction HR-HPV PCR was performed using the 23 HPV Genotyping Real-time PCR Kit (Hybribio, China) containing 17 high risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73 and 82, and 6 low risk HPV types: 6, 11, 42, 43, 44 and 81.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eInterpreting Results\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eRNAscope stained FFPE cell sections were scanned using Leica AT2 scanner (Leica, US). Whole sections were examined at 40\u0026times; magnification. RNAscope results of HPV were recorded based on signal location and positive cells. For probe-HPV-HR and probe-HPV-LR results, the signal locations of cells and the positive cell numbers in each sample, \u0026lt;3, ≧3≦10 or \u0026gt;10, were recorded. Besides classic RNAscope signals in cytoplasm or nuclear, there are HPV RNAs gathered as clusters above one or more cells, which were recorded as well. For TERC results, RNA signals in nucleus were recorded. Two gynecologists (Z. H. \u0026nbsp;and H. Y.) evaluated the scanned sections independently. If a disagreement occurred, they reviewed the case together and reached the final agreement. The interpretation was generally straightforward; therefore, no significant disagreements lead to incompatibility.\u0026nbsp;\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eIn current study, totally 115 patient cervical cell samples have been collected. 101 samples have been successfully prepared into FFPE blocks and passed RNAscope positive control tests using the probe of Hs-UBC. Three of 101 samples failed RNAscope negative control tests with background of probe DapB staining. All 98 QC passed samples were studied using RNAscope HPV high-risk probe (V-HPV-HR18) and HPV low-risk probe (V-HPV-LR6). Hs-TERC probe targeting on cell malignance has also been detected in the samples.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong 98 analyzed patient samples, 69 showed consistent results between RNAscope and PCR from patient cervical scraped cells. 46 of 69 patient samples were PCR and RNAscope both negative (supplemental table 1). 23 of 69 patient samples were PCR and RNAscope both positive of HPV high risk subtypes (table 1). Among them, 15 patients showed positive HPV RNAscope signals in cells (cytoplasm or nucleus) with or without RNA signal clusters located outside (above) of cells (figure 1, a-c), whereas another 8 samples only carried HPV RNA signals as clusters located out of cells (table 1) (figure 1, d-f). \u0026nbsp;Five patient samples exhibited lncRNA TERC signals in cell nucleus which indicated cell transformation signs (figure 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe rest 29 patient samples with inconsistent RNAscope and PCR results were further divided into 3 subgroups (table 2). The first subgroup included 8 patient samples which were HPV RNAscope positive, either high risk or low risk subtypes, however PCR HPV negative. In this subgroup, RNAscope results indicated 4 samples carried HPV RNA signals in cells (figure 3, a), whereas the other 4 carried HPV RNA signals as clusters above cells (table 2-1) (figure 3, b-e). Notably four samples in this subgroup came from patients with medical history of post-surgery of Loop Eelectrosurgical Excision Procedure (LEEP) or at Cervical Intraepithelial Neoplasia (CIN) I/II level, respectively. Moreover, all RNAscope positive signals limited in small number of cells (≦10) no matter signal exhibition phenotype (table 2-1).\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe 2\u003csup\u003end\u003c/sup\u003e subgroup included 9 samples which were PCR results positive however RNAscope negative (table 2-2). One patient sample exhibited TERC RNAscope signals with a history of CIN II and post-surgery of Leep for 3 years (figure 3, f).\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe last subgroup consisted of 12 patients which were HPV positive detected by both RNAscope and PCR methods, however inconsistent between high-risk and low-risk subtypes (table 2-3) (figure 4, a-e). Four patients had medical history of CKC treatment, laser CO2 vaporization therapy or CINIII, respectively. One patient (patient #48) in the subgroup had no previous HPV infection history, her sample exhibited HPV high-risk RNAscope signals in cytoplasm (\u0026gt;10) and HPV low risk RNAscope clusters. PCR results showed HPV 52 positive. Moreover, the patient TERC RNAscope signals were positive (figure 4, f).\u0026nbsp;\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003ePrevious studies have confirmed that p16 expression is associated with cervical lesion classification\u0026nbsp;[4], namely the heavier the degree of cervical lesions is, the higher the degree of p16 expresses. As a parameter to judge the disease severity, p16 indicates surgery necessity when the lesion has been developed to certain stages. Gynecologists would prefer a parameter which could predict the progression of cervical lesion, especially when the lesion is at the initial stage or even no lesions are there yet. In China, immunohistochemistry of p16 and ki67 are mostly performed on pathological samples which need invasive performance, \u003cem\u003ee.g.\u0026nbsp;\u003c/em\u003ecolposcopy, accompanied with bleeding and infections sometimes. For women during pregnancy, cervical colposcopy biopsy caused bleeding may stimulate contractions. It may increase the risk of miscarriage or premature delivery and increase the psychological burden during the gestation period. For no pregnancy patients, if the lesions are located at the cervical canal, commonly happened in postmenopausal women, it is hard to reach by colposcopy. In order to obtain the biopsy of those areas, cone cutting surgeries are commonly selected which will increase patient unnecessary injury burden, with the risk of missed diagnosis still. It is necessary to find a method to assess the disease severity without colposcopy and even predict the risk of cervical disease progression. The ideal assay results should be able to determine whether a colposcopy biopsy is unavoidable for patient with high-risk subtype of HPV infection.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHigh-risk subtypes of HPV infection are known to have chance to lead cervical cells into transforming stages and may have more chance to develop into high squamous intraepithelial lesion (HSIL)\u0026nbsp;[5]. HPV PCR tests are therefore widely performed in clinical to monitor cervical lesion. With the fact that HPV PCR results examine HPV DNAs without the information of viral activity status, there are HPV PCR positive samples carried no active virus from patients who may be overjudged for aggressive treatments or misestimated disease procession. Moreover, HPV infective cell numbers and the location information of HPV active virus in/near cervical cells are lacking by PCR tests. It is reasonable to estimate different disease progression if active HPV high risk viral RNAs have been detected widely spread in huge number of cells \u003cem\u003evs.\u0026nbsp;\u003c/em\u003eonly in several cells.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCurrent study is the first report trying to estimate cervical lesion progression by using RNAscope HPV \u003cem\u003ein situ\u003c/em\u003e hybridization in cervical scraped cells as we have known. By developing a new method which solved the main technique difficulties of cell detachment and RNA quality issue of cervical scraped cells, it makes RNA \u003cem\u003ein situ\u003c/em\u003e hybridization of HPV E6/E7 become feasible. The results uncovered a window to study active HPV infection status and how the viral locally interacts with cervical cells which provides observable clues for disease progression assessment. 98 patient samples which passed RNAscope QC have been examined by using HPV high-risk probes, RNAscope HPV low-risk probes and TERC probe which is a long non-coding RNA associated with high grade squamous intraepithelial neoplasia (HSIL) and progression of invasive carcinoma. RNAscope results have been compared to PCR assay data for further analysis. In most cases, cervical scraped cells were collected twice, the former set was used for PCR assays and the latter set was used for RNAscope assays. In several cases, PCR used cell samples were collected at different date based on patients\u0026rsquo; situation. The collection time variations may partly lead to the 29 inconsistence of HPV results between RNAscope and PCR assays. HPV DNA results detected by PCR study were compared to active HPV transcribed E6/E7 mRNAs captured by RNAscope assays, which may also lead to varied results if HPV virus infection happened previously however with inactive status.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRNAscope assay result demonstrated two types of HPV signals. One is typical RNA signal dots located in cell nuclear and cytoplasm. The other type of signals exhibited big clusters located above cells, which looks like \u0026ldquo;out of cell\u0026rdquo; signals. The latter signal phenotype has been observed in both high-risk and low-risk HPV probe detected results which demonstrate viral secreting status. In the patients whose sample were RNAscope positive whereas PCR negative, 4 were after-LEEP or CIN I-III stage. RNAscope positive signals in the samples were most out of cells, with only 4 patient samples showed HPV positive signals in nucleus (\u0026lt;10). In nine RNAscope negative whereas PCR positive samples, seven were HPV High-risk subtypes. RNAscope results negative may due to sample and tested cell variations, inactive virus status or other sample preparation caused unknown reasons. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFive of ninety-eight samples (patient 1, 5, 20, 26 and 48) exhibited active HPV E6/E7 mRNA signals in more than 10 cells (supplemental table 2). Moreover, patient 20 and 48 were TERC positive. Four (patient 1, 20, 26, 48) were high-risk HPV subtypes. Follow up records indicate that patient 1, 20 and 48 have received Leep (patient 1 and 20) and hysterectomy (patient 48), respectively, shortly after this study. To be noticed, patient 48 was detected HPV 52 positive by PCR tests with no infection history, whereas RNAscope discovered HPV high-risk subtype positive with more than 10 infective cells, plus TERC positive. Except Patent 20 and 48, there are five more patient (patient 40, 60, 61, 75 and 77) were TERC positive by RNA \u003cem\u003ein situ\u003c/em\u003e tests. Among them, patient 40, 60 and 77 carried HPV high- risk E6/E7 mRNA more than 3 cervical cells (supplemental table 2). Follow up records indicated that patient 40 and 77 had lost contact since 2019. Patient 60 follow-up records showed HPV low risk positive only (2020 May). Patient 61 follow-up records showed HPV PCR 52 positive (2020 June). To be noticed, patient 61 had received LEEP (2016. Oct) before the study starting. By 2019 Dec, patient 75 was both HPV PCR and TCT negative (supplemental table 2). \u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eCurrent results suggest that a combination of RNAscope positive signals of TERC and HPV high-risk subtype signals in more than 10 cells (cytoplasm or nucleus) may connect with cervical lesion fast progression which deserves highly attention.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eHuman papillomavirus (HPV)\u003c/p\u003e\n\u003cp\u003ePolymerase Chain Reaction (PCR)\u003c/p\u003e\n\u003cp\u003elong non-coding RNA (lncRNA)\u003c/p\u003e\n\u003cp\u003eInstitutional Review Board (IRB)\u003c/p\u003e\n\u003cp\u003eCommittee on Human Research in the Medical Sciences (CHRMS)\u003c/p\u003e\n\u003cp\u003eneutral buffered formalin (NBF)\u003c/p\u003e\n\u003cp\u003eroom temperature (RT)\u003c/p\u003e\n\u003cp\u003eFormalin-fixed paraffin-embedding (FFPE)\u003c/p\u003e\n\u003cp\u003ediaminopimelate B (DapB)\u003c/p\u003e\n\u003cp\u003ehuman ubiquitin C (Hs-UBC)\u003c/p\u003e\n\u003cp\u003equality control (QC)\u003c/p\u003e\n\u003cp\u003eLoop Eelectrosurgical Excision Procedure (LEEP)\u003c/p\u003e\n\u003cp\u003eCervical Intraepithelial Neoplasia (CIN)\u003c/p\u003e\n\u003cp\u003ehigh squamous intraepithelial lesion (HSIL)\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the scientists of Advanced Cell Diagnostics, Inc., Dr. Na Li and her team (Mr. Zhifu Zhang, Mrs. Ying Zhou, Mrs. Hongzhe Sun and Mrs. Xin Wang) for technical assistance.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor\u0026rsquo;s contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDr. Yu-Mei Wu designed the study and data analysis. Dr. Zhao Hui wrote the manuscript, performed patient sample collections, did RNA\u0026nbsp;\u003cem\u003ein situ\u003c/em\u003e experiments and data analysis. Dr. \u0026nbsp;Yue He, Bei Fan\u003csup\u003e\u0026nbsp;\u003c/sup\u003eand Yan Wang all participated patient sample collections and RNA\u003cem\u003e\u0026nbsp;in situ\u0026nbsp;\u003c/em\u003etests. Dr. Yue He did RNA \u003cem\u003ein situ\u0026nbsp;\u003c/em\u003edata scoring. \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support (grant no. ZYLX201705).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll detailed data and methods and material have been included in the material and method sections and result sections.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was conducted with the approval of the\u0026nbsp;Beijing Obstetrics and Gynecology hospital, Capital Medical University,\u0026nbsp;Institutional Review Board (IRB) Committee on Human Research in the Medical Sciences (CHRMS).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all individual participants included in the study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis article has no conflict of interest\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003e1.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Wardak S: \u003cstrong\u003eHuman Papillomavirus (HPV) and cervical cancer\u0026nbsp;\u003c/strong\u003e\u003cem\u003eMed Dosw Mikrobiol\u0026nbsp;\u003c/em\u003e2016, \u003cstrong\u003e68\u0026nbsp;\u003c/strong\u003e73-84.\u003c/p\u003e\n\u003cp\u003e2.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Major AL, Dvoř\u0026aacute;k V, Schwarzov\u0026aacute; J, Skřiv\u0026aacute;nek A, Mal\u0026iacute;k T, Pluta M, Mayboroda I, Grandjean EM: \u003cstrong\u003eEfficacy and safety of an adsorbent and anti-oxidative vaginal gel on CIN1 and 2, on high-risk HPV, and on p16/Ki-67: a randomized controlled trial.\u003c/strong\u003e \u003cem\u003eArch Gynecol Obstet\u0026nbsp;\u003c/em\u003e2020 Nov 20, \u003cstrong\u003edoi: 10.1007/s00404-020-05816-8\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e3.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Evans MF, Peng Z, Clark KM, Adamson CS-C, Ma X-J, Wu X, Wang H, Luo Y, Cooper K: \u003cstrong\u003eHPV E6/E7 RNA in situ hybridization signal patterns as biomarkers of three-tier cervical intraepithelial neoplasia grade.\u003c/strong\u003e \u003cem\u003ePLoS One\u0026nbsp;\u003c/em\u003e2014, \u003cstrong\u003e9:\u003c/strong\u003ee91142.\u003c/p\u003e\n\u003cp\u003e4.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Sano T, Oyama T, Kashiwabara K, Fukuda T, Nakajima T: \u003cstrong\u003eExpression Status of p16 Protein Is Associated with Human Papillomavirus Oncogenic Potential in Cervical and Genital Lesions.\u003c/strong\u003e \u003cem\u003eThe American Journal of Pathology\u0026nbsp;\u003c/em\u003e1998, \u003cstrong\u003e153,:\u003c/strong\u003e1741-1748.\u003c/p\u003e\n\u003cp\u003e5.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Regauer S, Reich O, Kashofer K: \u003cstrong\u003eThin variant of high-grade squamous intraepithelial lesion - relationship with high-risk and possibly carcinogenic human papilloma virus subtypes and somatic cancer gene mutations.\u003c/strong\u003e \u003cem\u003eHistopathology\u0026nbsp;\u003c/em\u003e2019 \u003cstrong\u003e75:\u003c/strong\u003e405-412.\u003c/p\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1 Comparison of RNAscope and PCR results in patient cervical scraped cells. The detection results are consistent between the two methods.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"51.30813953488372%\"\u003e\n \u003cp\u003eRNAscope results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"23.11046511627907%\"\u003e\n \u003cp\u003ePCR results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003epatient #\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eProbe V-HPV-HR18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eProbe V-HPV-LR6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.430232558139535%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eProbe Hs-TERC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003eHPV subtype\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003eAge\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003esample collector\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e16, 53, 56, 58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eASCUS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eHPV 53, 58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eCINII-III\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003epostsurgery of CKC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eLSIL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e16, 18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eCINI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003eNot test\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eEarly invasion in cervical carcinoma\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003epostsurgery of LEEP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eHPV infection history\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003enuclear, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e82, 42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e6, 59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecyto and nucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eCINI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003epostsurgery of LEEP 2 years\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eHPV infection history and CINI\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eCINI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eCINII\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eY\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.430232558139535%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e6, 51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"8.284883720930232%\"\u003e\n \u003cp\u003eP#71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.081395348837209%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.703488372093023%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.430232558139535%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.267441860465116%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.61046511627907%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.5%\"\u003e\n \u003cp\u003eCINI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.122093023255814%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.174418604651162%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTale 2-1 Comparison of RNAscope and PCR results in patient cervical scraped cells- RNAscope positive whereas PCR negative.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"49.598163030998855%\"\u003e\n \u003cp\u003eRNAscope results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"24.110218140068888%\"\u003e\n \u003cp\u003ePCR results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003epatient #\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eProbe V-HPV-HR18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eProbe V-HPV-LR6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eProbe Hs-TERC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eHPV subtype\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.991963260619976%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003eage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003esample collector\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003ecluster, \u0026gt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.991963260619976%\"\u003e\n \u003cp\u003epostsurgery of Loop Eelectrosurgical Excision Procedure (LEEP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003enucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.991963260619976%\"\u003e\n \u003cp\u003epostsurgery of Loop Eelectrosurgical Excision Procedure (LEEP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003enucleus, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003enucleus, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003ecluster, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003ecluster, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.991963260619976%\"\u003e\n \u003cp\u003eCINII，postsurgery of Loop Eelectrosurgical Excision Procedure(LEEP) 2 years\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003enucleus and cluster, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.991963260619976%\"\u003e\n \u003cp\u003eCINI Cervical Intraepithelial Neoplasia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.544202066590127%\"\u003e\n \u003cp\u003eP#80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12.399540757749714%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.332950631458095%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.7405281285878305%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.644087256027554%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.1182548794489096%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.991963260619976%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.495981630309988%\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.251435132032148%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2-2 Comparison of RNAscope and PCR results in patient cervical scraped cells- RNAscope negative whereas PCR positive.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"49.01960784313726%\"\u003e\n \u003cp\u003eRNAscope results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"23.414071510957324%\"\u003e\n \u003cp\u003ePCR results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003epatient #\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eProbe V-HPV-HR18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.649365628604382%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eProbe V-HPV-LR6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eProbe Hs-TERC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003eHPV subtype\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003eage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003esample collector\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.649365628604382%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e51, 52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003eCKC after\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e33, 52, 58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003enucleus, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003eCINI-II, postsurgery of \u0026nbsp; \u0026nbsp; Leep (3 years)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003eCINI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.57439446366782%\"\u003e\n \u003cp\u003eP#96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.649365628604382%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.767012687427912%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.380622837370241%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.151095732410611%\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"16.262975778546714%\"\u003e\n \u003cp\u003eVaINIII, Vaginal Intraepithelial Neoplasia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.688581314878892%\"\u003e\n \u003cp\u003e71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.303344867358708%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2-3 Comparison of RNAscope and PCR results in patient cervical scraped cells- RNAscope and PCR both positive whereas inconsistence between high-risk and low-risk subtypes.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"48.51936218678816%\"\u003e\n \u003cp\u003eRNAscope results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"32.46013667425968%\"\u003e\n \u003cp\u003ePCR results\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003epatient #\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eProbe V-HPV-HR18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eProbe V-HPV-LR6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eProbe Hs-TERC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003eDescription\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003eHPV subtype\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003eNote\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003eage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003esample collector\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003enuclear, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003enuclear, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e53, 56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003epostsurgery of \u0026nbsp;Cold Knife Conization(CKC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecyto, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e11, 59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e68, 43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e\u0026nbsp;postsurgery of \u0026nbsp;Cold Knife Conization(CKC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026gt;=3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003ecyto, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e44, 45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003eCINIII\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e16, 68, 42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003eHPV infection history, after laser CO2 vaporization therapy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003ecyto, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecluster, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003enucleus, \u0026gt;10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003ecyto, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"6.492027334851937%\"\u003e\n \u003cp\u003eP#73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.82004555808656%\"\u003e\n \u003cp\u003ecluster, \u0026lt;3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.694760820045558%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.136674259681094%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.922551252847381%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"10.250569476082005%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.061503416856492%\"\u003e\n \u003cp\u003e42, 43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.39863325740319%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"4.89749430523918%\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"7.6309794988610475%\"\u003e\n \u003cp\u003eDoctor #1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"virology-journal","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"virj","sideBox":"Learn more about [Virology Journal](http://virologyj.biomedcentral.com/)","snPcode":"12985","submissionUrl":"https://submission.nature.com/new-submission/12985/3","title":"Virology Journal","twitterHandle":"@VirologyJ","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"RNA in situ hybridization, HPV E6/E7, cervical scraped cells, cervical malignancy tests","lastPublishedDoi":"10.21203/rs.3.rs-838435/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-838435/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground\u003c/p\u003e\u003cp\u003eHuman papillomavirus screen in female cervical cells has demonstrated values in clinical diagnosis of precancerous lesions and cervical cancers. Human papillomavirus tests of cervical cells by utilizing Polymerase Chain Reaction (PCR) method provides human papillomavirus infection status however no further virus \u003cem\u003ein situ\u003c/em\u003e information. Although it is well known that the tests of human papillomavirus E6/E7 RNA location in infected cervical cells and cell internal malignancy molecular will provide clues for gynecologists to evaluate disease progression, there are technique difficulties to preserve RNAs in cervical scraped cells for \u003cem\u003ein situ\u003c/em\u003e hybridization. \u003c/p\u003e\u003cp\u003eMethods\u003c/p\u003e\u003cp\u003eIn current study, after developing a cervical cell collection and preparation methods for RNA\u003cem\u003e in situ\u003c/em\u003e hybridization, we captured the chance to screen 98 patient cervical cell samples and detected human papillomavirus E6/E7 mRNAs of high-risk subtypes, low-risk subtypes and lncRNA TERC in the cells. \u003c/p\u003e\u003cp\u003eResults\u003c/p\u003e\u003cp\u003eThere are 70% consistence between human papillomavirus PCR and human papillomavirus RNA \u003cem\u003ein situ\u003c/em\u003e hybridization results in cervical collected cells. Viral E6/E7 mRNAs were observed to distribute in cervical cell nuclear and cytoplasm. Moreover, viral gathered clusters were observed outside of cells through human papillomavirus RNA \u003cem\u003ein situ \u003c/em\u003ehybridization. Varied numbers of human papillomavirus infective cells were detected by RNAscope assay in different patients even though they are all human papillomavirus high-risk subtype positive discovered by human papillomavirus PCR results. A cell malignancy related long non-coding RNA, TERC, has been detected in seven patient samples. The patient follow-up information was further analyzed with RNAscope results which indicated a combination of RNAscope positive signals of TERC and human papillomavirus high risk signals in more than 10 cells (cytoplasm or nucleus) may connect with cervical lesion fast progression which deserves further studies in the future.\u003c/p\u003e\u003cp\u003eConclusions\u003c/p\u003e\u003cp\u003eTaken together, current study has provided an observable clue for gynecologists to evaluate human papillomavirus infection stage and cell malignancy status which may contribute for assessment of cervical disease progression.\u0026nbsp;\u003c/p\u003e","manuscriptTitle":"Human Papillomavirus E6E7 mRNA and TERC lncRNA in Situ Detection in Cervical Scraped Cells and Cervical Disease Progression Assessment","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-09-13 15:24:18","doi":"10.21203/rs.3.rs-838435/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-10-04T00:00:00+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-10-03T00:00:00+00:00","index":3,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"editorInvitedReview","content":"","date":"2021-10-02T00:00:00+00:00","index":2,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"editorInvitedReview","content":"","date":"2021-09-27T00:00:00+00:00","index":1,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"reviewerAgreed","content":"","date":"2021-09-19T00:00:00+00:00","index":3,"fulltext":""},{"type":"reviewerAgreed","content":"","date":"2021-09-18T00:00:00+00:00","index":2,"fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-09-17T12:17:19+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-09-17T12:04:15+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2021-09-17T00:00:00+00:00","index":1,"fulltext":""},{"type":"checksComplete","content":"","date":"2021-09-06T23:00:00+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Virology Journal","date":"2021-09-03T11:21:46+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-09-03T04:14:35+00:00","index":"","fulltext":""},{"type":"submitted","content":"Virology Journal","date":"2021-08-22T22:27:30+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"virology-journal","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"virj","sideBox":"Learn more about [Virology Journal](http://virologyj.biomedcentral.com/)","snPcode":"12985","submissionUrl":"https://submission.nature.com/new-submission/12985/3","title":"Virology Journal","twitterHandle":"@VirologyJ","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2006392e-b2de-474e-a421-ce5be5d20578","owner":[],"postedDate":"September 13th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":7134506,"name":"Virology"}],"tags":[],"updatedAt":"2022-01-24T10:51:46+00:00","versionOfRecord":{"articleIdentity":"rs-838435","link":"https://doi.org/10.1186/s12985-021-01696-9","journal":{"identity":"virology-journal","isVorOnly":false,"title":"Virology Journal"},"publishedOn":"2022-01-24 10:51:46","publishedOnDateReadable":"January 24th, 2022"},"versionCreatedAt":"2021-09-13 15:24:18","video":"","vorDoi":"10.1186/s12985-021-01696-9","vorDoiUrl":"https://doi.org/10.1186/s12985-021-01696-9","workflowStages":[]},"version":"v1","identity":"rs-838435","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-838435","identity":"rs-838435","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}