Identification of candidate microRNA markers of endometriosis with the use of next-generation sequencing and quantitative real-time polymerase chain reaction

Elahe Papari, Papari E, Mehrdad Noruzinia, Ladan Kashani, Kashani L, Foster WG, Warren G Foster
Fertility and sterility · 2020 · vol. 113(6) , pp. 1232–1241 · doi:10.1016/j.fertnstert.2020.01.026 · PMID:32482255 · W3029129472
article OA: closed CC0 ⤵ 45 in-corpus citations
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This study used next-generation sequencing and qPCR to identify microRNAs that could serve as diagnostic markers for endometriosis.

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Abstract

ObjectiveTo identify novel candidate diagnostic microRNA (miRNA) markers of endometriosis by means of an unbiased search with confirmation by means of targeted polymerase chain reaction (PCR).DesignRetrospective cohort.SettingUniversity teaching hospitals.Patient(s)Women with endometriosis and control women, confirmed with the use of laparoscopy.Interventions(s)Diagnostic laparoscopy and blood sample.Main outcome measure(s)Next-generation sequencing (NGS) and quantitative real-time PCR (qRT-PCR).Result(s)Candidate miRNAs differentially expressed in women with endometriosis compared with control women were identified by means of NGS and selected for qRT-PCR. Plasma samples from another cohort of women with surgically confirmed endometriosis (n = 53) and disease-free control women (n = 53) were checked for hemolysis using spectrophotometry and the ratio of miR-23a and miR-451 by means of qRT-PCR. MicroRNA signatures were quantified by means of qRT-PCR in hemolysis-free plasma samples of case subjects (n = 25) and control subjects (n = 28) with the use of miRcury LNA miRNA. Circulating levels of eight miRNAs (miR-199a-3p, miR-143-3p, miR-340-5p, let-7b-5p, miR-21-5p, miR-17-5p, miR-20a-5p, and miR-103a-3p) were significantly lower in case subjects compared to control subjects. The sensitivity and specificity for individual miRNAs ranged from 0.36 to 1.00 and from 0.43 to 1.00, respectively, but when combined produced sensitivity and specificity of 0.92 and 0.86 with positive (PPV) and (NPV) predictive values of 0.85 and 0.92, respectively. However, combination of five miRNAs (miR-17-5p, miR-20a-5p, miR-199a-3p, miR-143-3p, and let-7b-5p) produced sensitivity and specificity of 0.96 and 0.79 with PPV and NPV of 0.80 and 0.96, respectively.Conclusion(s)We conclude that a panel of candidate miRNAs was comparable to laparoscopy in distinguishing between women with endometriosis and control women.

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Condition tags

endometriosis

MeSH descriptors

Circulating MicroRNA Endometriosis High-Throughput Nucleotide Sequencing Real-Time Polymerase Chain Reaction Adult Biomarkers Biomarkers Circulating MicroRNA Circulating MicroRNA Endometriosis Endometriosis Endometriosis Female Humans Laparoscopy Middle Aged Predictive Value of Tests Reproducibility of Results Retrospective Studies Young Adult

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