Evaluation of the Performance of a Biofire FilmArray Meningitis/Encephalitis Multiplex Real-Time PCR Assay for Bacterial Meningitis Diagnosis at a Tertiary Care Hospital

Evaluation of the Performance of a Biofire FilmArray Meningitis/Encephalitis Multiplex Real-Time PCR Assay for Bacterial Meningitis Diagnosis at a Tertiary Care Hospital | 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 Article Evaluation of the Performance of a Biofire FilmArray Meningitis/Encephalitis Multiplex Real-Time PCR Assay for Bacterial Meningitis Diagnosis at a Tertiary Care Hospital Shabnam Dildar, Saman Nadeem, Nayab Afzal, Muhammad Ammar Athar This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5184548/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Meningitis and encephalitis are critical, life-threatening conditions associated with high rates of morbidity and mortality. Early diagnosis and immediate treatment are essential for decreasing mortality and improving patient outcomes. The Biofire Film Array Meningitis/Encephalitis (ME) panel is a multiplex polymerase chain reaction (PCR)-nucleic acid-based diagnostic assay designed for the rapid diagnosis of bacterial, viral, and fungal pathogens in cerebrospinal fluid. In this study, we evaluated the performance of Biofire Film Array Meningitis/Encephalitis multiplexing real a biofire film array meningitis/encephalitis multiplex real-time PCR assay for the detection of bacterial pathogens in suspected cases of bacterial meningitis. Methods: Cerebrospinal fluid (CSF) and blood samples were collected from 132 intensive care unit (ICU) patients with suspected bacterial meningitis. The clinical samples were analyzed using traditional culture and sensitivity methods, the Biofire Film Array ME Panel multiplex PCR, procalcitonin (PCT), and C-reactive protein (CRP) assays. The sensitivity, specificity and diagnostic accuracy of the Biofire Film Array ME Panel real-time PCR assay were evaluated. Results: A total of 132 patients, with a mean age of 38.9 ± 23.7 years, were included in the study. The mean levels of C-reactive protein (CRP) and procalcitonin (PCT) were 52.41 ± 82.23 mg/L and 3.7 ± 7.6 ng/mL, respectively. Among these patients, 9 (6.8%) had positive cultures for Listeria monocytogenes (n = 6), Streptococcus pneumonia (n = 2), and Neisseria meningitides (n = 1), while 123 patients were culture negative. All 9 culture-positive cases were detected by the Biofire ME Panel, and 114 out of 123 (92.7%) culture-negative cases were confirmed as true negatives by the panel. The Biofire ME Panel demonstrated excellent diagnostic performance, with a sensitivity of 100% (95% CI: 63.06–100%), a specificity of 91.94% (95% CI: 85.67–96.06%), and an overall diagnostic accuracy of 92.42% (95% CI: 86.51–96.31%). Conclusion : The Biofire ME Panel is strongly correlatedwith CSF culture and has excellent diagnostic accuracy for bacterial meningitis. Moreover, the NRS-2002 may be used as a routine test for suspected cases of bacterial meningitis for early diagnosis and optimal treatment to reduce mortality and morbidity. Biofire FilmArray ME Panel Real-Time PCR Bacterial Meningitis CSF Culture Introduction Infections of the central nervous system (CNS) are common causes of morbidity and mortality. Accurate and rapid diagnosis is essential for effective management. Meningitis is a serious and potentially life-threatening condition involving inflammation of the protective membranes (meanings) covering the brain and spinal cord. It is commonly caused by bacterial, viral, or fungal infections (1). Traditional diagnostic techniques, such as blood culture, are considered the standard methods for identifying bacterial pathogens. However, these methods often suffer from long processing times and decreased sensitivity, especially after antibiotic administration. Such delays in diagnosis can significantly impact patient morbidity and mortality rates, highlighting the need for more efficient diagnostic alternatives (2, 3). Recent advancements in molecular diagnostics, including multiplex real-time PCR melting curve analysis panel screening, have improved the ability to simultaneously detect multiple targeted pathogens and have opened up new possibilities and challenges (4). The Biofire FilmArray ME Panel is a multiplex real-time PCR assay (ME PCR Panel; BioFire Diagnostics, Salt Lake City, Utah) designed to rapidly detect a comprehensive range of 14 pathogens responsible for meningitis, including bacteria, viruses, and fungi. This high-throughput molecular diagnostic tool has shown promising results in terms of sensitivity and specificity in detecting a wide array of pathogens associated with meningitis. The Film Array® Meningitis/Encephalitis (ME) panel was approved by the Food and Drug Administration (FDA) in October 2015. This panel was developed by Biofire Diagnostics in Salt Lake City, UT, and represents the first FDA-approved multiplex real-time PCR assay designed for the evaluation of cerebrospinal fluid (CSF) samples. According to a recent meta-analysis, the Film Array ME Panel demonstrated a sensitivity of 90% and a specificity of 97% for detecting various microorganisms. Specifically, the sensitivity varied from 85.7% for HHV-6 to 100% for nine of the fourteen targeted organisms. However, it is important to note that the study had limited samples for many organisms and did not detect L. monocytogenes or N. meningitidis . The panel’s specificity is notably high at 99.2% (5, 6, 7). Compared with traditional CSF cultures, the Biofire ME Panel has a rapid turnaround time, which may take several days to produce results. A minimum of 0.2 mL of CSF is needed, and 12 samples can be processed simultaneously. The most essential feature is the ability to obtain results in approximately one hour, significantly accelerating the diagnostic process. Early diagnosis and initiation of treatment are crucial in patients with meningitis, and early intervention can significantly impact patient outcomes (5). To thoroughly evaluate the diagnostic accuracy of the Biofire ME Panel, extensive clinical studies and comparative analyses with conventional diagnostic methods are necessary. These studies can provide a deeper understanding of the test's performance in various clinical scenarios, ultimately enhancing its clinical utility and impact on patient outcomes. This study aimed to assess the diagnostic accuracy of the Biofire Film Array ME Panel compared to traditional CSF culture methods. By examining the sensitivity, specificity, and overall diagnostic effectiveness of the Biofire Film Array test, we intend to establish its role as a primary diagnostic tool for bacterial meningitis. Methods This cross-sectional study was conducted in the laboratory of National Medical Centre (NMC) Hospital, Karachi, Pakistan, from January 2023 to December 2023. All patients admitted to the Medical Intensive Care Unit (ICU) with suspected meningitis were included in the study. Suspected cases of bacterial meningitis were defined according to WHO guidelines. Approval for the study was obtained from the Institutional Review Board (IRB) with reference number NMC/ERC/001/2024 prior to commencement. Procalcitonin (ProCT) levels were analyzed using the electrochemiluminescence technique on the Roche Cobas e-411 analyzer, with cutoff values established by the Consensus Conference of the American College of Chest Physicians/Society of Critical Care Medicine. The ProCT levels were categorized as follows: <0.5 ng/ml indicated a low risk of sepsis, ≥0.5 ng/ml but <2 ng/ml indicated moderate risk, ≥2 ng/ml but < 10 ng/ml indicated high risk, and ≥ 10 ng/ml indicated severe sepsis or septic shock [6-8]. C-reactive protein (CRP) levels were analyzed using the particle-enhanced immunoturbidimetric assay on the Roche Cobas c-311 analyzer, with a cutoff value of CRP <10 mg/L. Cerebrospinal fluid (CSF) culture samples were inoculated on chocolate agar, MacConkey agar, and 5% sheep blood agar media, followed by Gram staining. The plates were incubated in an ESCO Cel Culture® CO₂ incubator at 37°C and 5% CO₂ for 24-48 hours. The plates were assessed visually for growth, and Gram staining was performed. The samples were categorized as gram-positive or gram-negative bacteria. Specific microorganism identification and antibiotic susceptibility testing were conducted using the Vitek 2 Compact (BioMérieux) system CSF (200 µL) and the hydration solution were added to a Biofire® FilmArray® ME Panel pouch. The prepared pouch was subsequently inserted into the Biofire® FilmArray® PCR automated machine for a one-hour processing cycle. The system automatically extracts and purifies nucleic acids from the lysed sample. Multiplex real-time PCR was conducted in two stages. The first stage involves a high-capacity, massively multiplexed reaction (PCR1), followed by multiple second-stage PCRs (PCR2), which amplify specific sequences from the PCR1 products. The Biofire® FilmArray® real-time PCR system utilizes endpoint-melting curve analysis to identify and generate results for each of the 14 target pathogens represented in the Biofire ME Panel array. The results were automatically interpreted by Biofire® Film Array® software. All procedures were performed in strict accordance with the manufacturer's instructions to ensure accuracy and reliability. All the collected data were analyzed using SPSS version 19. The results are expressed as the mean ± standard deviation, median, and interquartile range for variables such as CRP and PCT. Frequencies and percentages were computed for categorical variables, including sex, CSF culture results, and Biofire ME Panel outcomes. The performance of the Biofire ME Panel was evaluated in terms of sensitivity, specificity, and diagnostic accuracy. Results A total of 132 ICU patients were included in the study, and cerebrospinal fluid (CSF) and blood samples were collected for analysis. Among these patients, 58 (43.9%) were female, and 74 (56.1%) were male. The median age of the patients in the cohort was 38.5 years (IQR: 23–55). The median C-reactive protein (CRP) level was 3.7 mg/L (IQR: 1.85–11.75), and the median procalcitonin (PCT) level was 0.10 ng/mL (IQR: 0.025–0.40). Of the 132 patients, 9 (6.8%) had positive cultures for Listeria monocytogenes (n = 6), Streptococcus pneumoniae (n = 2), and Neisseria meningitidis (n = 1), while 123 patients were culture negative. All 9 culture-positive cases were detected by the Biofire ME Panel, and 114 out of 123 (92.7%) culture-negative cases were confirmed to be true negatives by the Biofire ME Panel. However, among the 10 false-positive patients, 7 (70%) were incorrectly identified as Streptococcus. The average time to detect organisms using the Biofire ME Panel assay was 3 ± 1 hour after sample collection, whereas CSF culture provided specific microorganism identification and antibiotic susceptibility results within 48 ± 2 hours. In patients with positive CSF cultures, the mean CRP and PCT levels were significantly elevated, at 61.23 ± 20.38 mg/L and 34.47 ± 22.78 ng/mL, respectively. In contrast, patients with negative CSF cultures had markedly lower mean C-reactive protein (CRP) and thrombocytocrit (PCT) levels, recorded at 4.73 ± 4.27 mg/L and 0.41 ± 1.90 ng/mL, respectively. The Biofire ME Panel demonstrated strong performance in diagnosing meningitis, with a sensitivity of 100% (95% CI: 63.06–100%), a specificity of 91.94% (95% CI: 85.67–96.06%), and an overall diagnostic accuracy of 92.42% (95% CI: 86.51–96.31%), as shown in Table numbe 1. Table 1 Diagnostic accuracy of the Biofire ME Real-Time PCR Assay Compared to Culture n = 132 CSF Culture Positive Negative Detected 8(TP) 10(FP) Not Detected 0(FN) 114(TN) Total 8 124 Biofire ME Real Time PCR Assay Sensitivity 8/8 100% Specificity 114/124 91.94% Diagnostic accuracy 8 + 114/132 92.42% * Sensitivity = TP/(TP + FN); specificity = TN/(TN + FP); diagnostic accuracy=(TP + TN)/(TP + TN + FP + FN) Discussion Infectious meningitis is a serious condition characterized by signs and symptoms such as high-grade fever, nausea, altered consciousness and neck stiffness. These signs can be nonspecific and may overlap with symptoms of other illnesses. While viral meningitis is also common, it is most rapidly fatal ( 8 ). Therefore, rapid diagnosis is essential for proper treatment and patient survival. Conventional diagnosis requires laboratory tests such as CSF detailed reports, CSF Gram staining and other biomarkers (PCT, CRP, etc.). However, biomarkers lack specificity, while CSF cultures take several days to obtain results. Recent advancements in the field of molecular diagnostics, especially PCR, are revolutionizing infectious disease management in acute-care settings with rapid, accurate, and potentially cost-effective tools for pathogen detection and antimicrobial resistance profiling ( 9 , 10 )(reference Ammar review). For the diagnosis of meningitis, the rapid diagnostic test Biofire® FilmArray® ME Panel has the capacity to detect 14 different pathogen panels simultaneously based on multiplexed/meclting curve analysis real-time PCR technology and provides rapid results ( 6 ). There was a slight male predominance in the cohort studied, with 56.1% males. The median age was 38.50 years, which provided insight into the age distribution of the individuals studied. The median C-reactive protein level was 3.7 mg/L, and the median procalcitonin level was 0.100 ng/mL. In patients in whom CSF culture was positive, the mean CRP and PCT values were substantially greater, indicating that these biomarkers are elevated in patients with confirmed meningitis and suggesting their potential utility in the diagnosis of this disease. This finding is consistent with the expectation that patients with active infections (e.g., meningitis) would have elevated inflammatory marker levels. Similar findings have been reported elsewhere ( 11 ). The study showed that 8 (6.06%) patients exhibited positive CSF culture growth, indicating bacterial meningitis. Listeria monocytogenes was the most commonly isolated organism, whereas previous studies have identified Streptococcus pneumoniae as the most common pathogen [12–13], likely due to differences in climate, hygiene, and population density. The BioFire® real-time PCR ME Panel detected all 8 culture-positive cases, with a sensitivity of 100%. This exceptional sensitivity highlights the panel’s efficacy in accurately identifying true-positive cases, thereby ensuring that no cases of bacterial meningitis were missed. Similar findings of high sensitivity were reported by Piccirilli et al. and Lee et al. in their respective studies ( 12 , 13 ). The BioFire® ME Panel demonstrated a specificity of 91.94%, accurately identifying 114 true negatives out of 124 culture-negative specimens, consistent with findings from other studies ( 14 )( 15 ), although Ra`dmard et al. reported a lower specificity of 85.7% ( 15 )[17]. Importantly, no false negatives were observed, highlighting the strength of the BioFire® ME panel in detecting true-positive cases. However, 10 false positives were identified, indicating that the BioFire® ME panel assay may occasionally detect pathogens where none exist. This suggests the need for confirmatory tests or additional clinical evaluation to prevent unnecessary treatment or invasive procedures, a concern similarly noted by Hansen et al. regarding false positives in their study ( 14 ). Despite its high sensitivity and specificity, clinicians should be cautious of the potential harm that false positives can cause, including unnecessary or potentially toxic treatments. This panel screening assay achieved an overall diagnostic accuracy of 92.42%, which combined both sensitivity and specificity. Therefore, this level of accuracy indicates that the panel may have an important role in the diagnosis and management of meningitis within appropriate time limits. CSF culture is still considered the gold standard for diagnosing meningitis; however, this method is typically slow and may miss some bacteria. In contrast, the BioFire® ME Panel provides not only excellent sensitivity and specificity but also a quick turnaround time, providing significant benefit in clinical settings by allowing timely and precise diagnosis. However, despite its excellent performance, the Biofire® ME screening panel has certain limitations. A false positive rate (10/132) could lead to unnecessary treatments and follow-up tests. Furthermore, while CSF culture is regarded as the gold standard, it may not detect all pathogens; therefore, the true performance of the Biofire ME Panel screening assay could vary depending on the actual pathogen distribution across patient groups. Our study has several limitations. When cerebrospinal fluid (CSF) culture is used as the gold standard, culture methods can yield false negatives, potentially impacting the perceived accuracy of the BioFire® ME Panel. Although the results are promising, further research with larger sample sizes and more diverse populations is necessary to validate the panel’s performance across different geographic regions and patient demographics. Additionally, the limited number of samples may have contributed to the absence of certain organisms in the study. Furthermore, this study did not assess how the BioFire® ME Panel results correlated with clinical findings, leaving an important aspect of diagnostic utility unexplored. Conclusion In conclusion, our study suggested that the Biofire ME Panel is a valuable tool for diagnosing meningitis, given its high sensitivity, specificity, and overall diagnostic accuracy. This rapid and comprehensive diagnostic method could facilitate prompt and appropriate treatment, ultimately improving patient outcomes. Declarations Authors’ contributions All authors have read and approved the manuscript. SD: wrote the main text of manuscript and collect data SN: wrote manuscript and analyze data NA: wrote manuscript and analyze data MAA:critically reviewed the manuscript for intellectual content. Funding None. Availability of data and materials All relevant data are included in the manuscript. Ethics approval and consent to participate The approval of Institutional Review Board (IRB)/Ethical Committee of NMC Hospital was taken for study. Consent for publication Written informed consent was obtained from the all patients. Competing interests The authors declare that they have no competing interests. Dedication: To my Mother (Late Zarina Ramzan Ali), Father (Late Ramzan Ali) and Brother (Late Rehman Ali). ACKNOWLEDGEMENTS : None. References Trujillo-Gómez J, Tsokani S, Arango-Ferreira C, Atehortúa-Muñoz S, Jimenez-Villegas MJ, Serrano-Tabares C, et al. Biofire FilmArray Meningitis/Encephalitis panel for the aetiological diagnosis of central nervous system infections: A systematic review and diagnostic test accuracy meta-analysis. EClinicalMedicine. 2022;44:101275. Brouwer MC, Tunkel AR, van de Beek D. Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis. Clin Microbiol Rev. 2010;23(3):467–92. Leber AL, Everhart K, Balada-Llasat JM, Cullison J, Daly J, Holt S, et al. Multicenter Evaluation of BioFire FilmArray Meningitis/Encephalitis Panel for Detection of Bacteria, Viruses, and Yeast in Cerebrospinal Fluid Specimens. J Clin Microbiol. 2016;54(9):2251–61. Athar MA, Xu Y, Xie X, Xu Z, Ahmad V, Hayder Z, et al. Rapid detection of HCV genotyping 1a, 1b, 2a, 3a, 3b and 6a in a single reaction using two-melting temperature codes by a real-time PCR-based assay. J Virol Methods. 2015;222:85–90. Ramanan P, Bryson AL, Binnicker MJ, Pritt BS, Patel R. Syndromic Panel-Based Testing in Clinical Microbiology. Clin Microbiol Rev. 2018;31(1). Hanson KE. The First Fully Automated Molecular Diagnostic Panel for Meningitis and Encephalitis: How Well Does It Perform, and When Should It Be Used? J Clin Microbiol. 2016;54(9):2222–4. Tansarli GS, Chapin KC. Diagnostic test accuracy of the BioFire® FilmArray® meningitis/encephalitis panel: a systematic review and meta-analysis. Clin Microbiol Infect. 2020;26(3):281–90. He T, Kaplan S, Kamboj M, Tang YW. Laboratory Diagnosis of Central Nervous System Infection. Curr Infect Dis Rep. 2016;18(11):35. Yang S, Rothman RE. PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings. Lancet Infect Dis. 2004;4(6):337–48. Ullah MAAVAIUS. Molecular Diagnosis of Hepatitis C Viruses; Technologies and Their Clinical Applications. Microbiol Immunological Commun. 2022;1(1):55–73. Sager R, Kutz A, Mueller B, Schuetz P. Procalcitonin-guided diagnosis and antibiotic stewardship revisited. BMC Med. 2017;15(1):15. Ali SA, Taj MK, Ali SH. Antimicrobial Resistance Pattern of Bacterial Meningitis Among Patients in Quetta, Pakistan. Infect Drug Resist. 2021;14:5107–20. Lee SH, Chen SY, Chien JY, Lee TF, Chen JM, Hsueh PR. Usefulness of the FilmArray meningitis/encephalitis (M/E) panel for the diagnosis of infectious meningitis and encephalitis in Taiwan. J Microbiol Immunol Infect. 2019;52(5):760–8. Hanson KE, Slechta ES, Killpack JA, Heyrend C, Lunt T, Daly JA, et al. Preclinical Assessment of a Fully Automated Multiplex PCR Panel for Detection of Central Nervous System Pathogens. J Clin Microbiol. 2016;54(3):785–7. Messacar K, Breazeale G, Robinson CC, Dominguez SR. Potential clinical impact of the film array meningitis encephalitis panel in children with suspected central nervous system infections. Diagn Microbiol Infect Dis. 2016;86(1):118–20. Additional Declarations The authors declare no competing interests. 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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-5184548","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":362175942,"identity":"2febbf2a-24cf-4740-a768-a09f7d3ac5fe","order_by":0,"name":"Shabnam Dildar","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIiWNgGAWjYDACCQY2COMwc4MEA4MNSIRoLYwgLWmkaDkA1nKYsBb52c3PHhfUbGPgO87YeOPnnvOJ/bObDz5gqLGJxqXF4M4xc+MZx24zSB5mbLbseXY7ccadY8kGDMfSchtwaZFIMJPmYbvNYHCYsU2C58DtxIYbOWYSjA2HcWqRn5H+TZrnH0SL5J8D5xLnE9LCAFQgzdsG0SLNc+BA4gZCWgxu5JQb8/bd5gH5xVrmQLLxxhtpyQYJePwCdNi2xzzfbsvxnT988OabA3ay824kH3zwocYGt8OggAfGcASrTCCgHAXYk6J4FIyCUTAKRgYAABiWYJ81l9zUAAAAAElFTkSuQmCC","orcid":"","institution":"National Medical Centre(NMC)","correspondingAuthor":true,"prefix":"","firstName":"Shabnam","middleName":"","lastName":"Dildar","suffix":""},{"id":362175943,"identity":"f1e414d4-1468-4d5f-a27f-126c32a1b038","order_by":1,"name":"Saman Nadeem","email":"","orcid":"","institution":"National Medical Centre(NMC)","correspondingAuthor":false,"prefix":"","firstName":"Saman","middleName":"","lastName":"Nadeem","suffix":""},{"id":362175944,"identity":"56a0b0f5-1c33-4cf6-8d5b-8da1cc012cb4","order_by":2,"name":"Nayab Afzal","email":"","orcid":"","institution":"Aga Khan University Hospital","correspondingAuthor":false,"prefix":"","firstName":"Nayab","middleName":"","lastName":"Afzal","suffix":""},{"id":362175945,"identity":"2f0677d1-f06e-40a6-825c-5de9c960b352","order_by":3,"name":"Muhammad Ammar Athar","email":"","orcid":"","institution":"National Medical Centre(NMC)","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"Ammar","lastName":"Athar","suffix":""}],"badges":[],"createdAt":"2024-10-01 04:53:19","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-5184548/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5184548/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":66162138,"identity":"5aaea636-df89-4ab5-8a43-73a4e5dfd0e2","added_by":"auto","created_at":"2024-10-08 09:25:49","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":321860,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5184548/v1/65b9a062-ab13-4def-a574-799dc541ecb6.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"Evaluation of the Performance of a Biofire FilmArray Meningitis/Encephalitis Multiplex Real-Time PCR Assay for Bacterial Meningitis Diagnosis at a Tertiary Care Hospital","fulltext":[{"header":"Introduction","content":"\u003cp\u003eInfections\u0026nbsp;of the central nervous system (CNS) are common\u0026nbsp;causes\u0026nbsp;of morbidity and mortality. Accurate and rapid diagnosis is essential for effective management. Meningitis is a serious and potentially life-threatening condition\u0026nbsp;involving\u0026nbsp;inflammation of the protective membranes (meanings) covering the brain and spinal cord. It is commonly caused by bacterial, viral, or fungal infections (1).\u0026nbsp;Traditional diagnostic techniques, such as blood\u0026nbsp;culture, are considered the standard\u0026nbsp;methods\u0026nbsp;for identifying bacterial pathogens. However, these methods often suffer from long processing times and decreased sensitivity, especially after antibiotic administration. Such delays in diagnosis can significantly impact patient morbidity and mortality rates, highlighting the need for more efficient diagnostic alternatives (2, 3).\u003c/p\u003e\n\u003cp\u003eRecent advancements in molecular diagnostics, including multiplex real-time PCR melting curve analysis panel screening, have improved the ability to simultaneously detect multiple targeted pathogens and have opened up new possibilities and challenges (4). The Biofire FilmArray ME Panel is a multiplex real-time PCR assay (ME PCR Panel; BioFire Diagnostics, Salt Lake City, Utah) designed to rapidly detect a comprehensive range of 14 pathogens responsible for meningitis, including bacteria, viruses, and fungi. This high-throughput molecular diagnostic tool has shown promising results in terms of sensitivity and specificity in detecting a wide array of pathogens associated with meningitis.\u003c/p\u003e\n\u003cp\u003eThe Film Array\u0026reg; Meningitis/Encephalitis (ME) panel was approved by the Food and Drug Administration (FDA) in October 2015. This panel was developed by Biofire Diagnostics in Salt Lake City, UT, and represents the first FDA-approved multiplex real-time PCR assay designed for the evaluation of cerebrospinal fluid (CSF) samples. According to a recent meta-analysis, the Film Array ME Panel demonstrated a sensitivity of 90% and a specificity of 97% for detecting various microorganisms. Specifically, the sensitivity varied from 85.7% for HHV-6 to 100% for nine of the fourteen targeted organisms. However, it is important to note that the study had limited samples for many organisms and did not detect \u003cem\u003eL. monocytogenes\u003c/em\u003e or \u003cem\u003eN. meningitidis\u003c/em\u003e. The panel\u0026rsquo;s specificity is notably high at 99.2% (5, 6, 7).\u003c/p\u003e\n\u003cp\u003eCompared with traditional CSF cultures, the Biofire ME Panel has a rapid turnaround time, which may take several days to produce results. A minimum of 0.2 mL of CSF is needed, and 12 samples can be processed simultaneously. The most essential feature is the ability to obtain results in approximately one hour, significantly accelerating the diagnostic process. Early diagnosis and initiation of treatment are crucial in patients with meningitis, and early intervention can significantly impact patient outcomes (5).\u003c/p\u003e\n\u003cp\u003eTo thoroughly evaluate the diagnostic accuracy of the Biofire ME Panel, extensive clinical studies and comparative analyses with conventional diagnostic methods are necessary. These studies can provide a deeper understanding of the test\u0026apos;s performance in various clinical scenarios, ultimately enhancing its clinical utility and impact on patient outcomes.\u003c/p\u003e\n\u003cp\u003eThis study aimed to assess the diagnostic accuracy of the Biofire Film Array ME Panel compared to traditional CSF culture methods. By examining the sensitivity, specificity, and overall diagnostic effectiveness of the Biofire Film Array test, we intend to establish its role as a primary diagnostic tool for bacterial meningitis.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eThis cross-sectional study was conducted in the laboratory of National Medical Centre (NMC) Hospital, Karachi, Pakistan, from January 2023 to December 2023. All patients admitted to the Medical Intensive Care Unit (ICU) with suspected meningitis were included in the study. Suspected cases of bacterial meningitis were defined according to WHO guidelines. Approval for the study was obtained from the Institutional Review Board (IRB) with reference number NMC/ERC/001/2024 prior to commencement.\u003c/p\u003e\n\u003cp\u003eProcalcitonin (ProCT) levels were analyzed using the electrochemiluminescence technique on the Roche Cobas e-411 analyzer, with cutoff values established by the Consensus Conference of the American College of Chest Physicians/Society of Critical Care Medicine. The ProCT levels were categorized as follows: \u0026lt;0.5 ng/ml indicated a low risk of sepsis, \u0026ge;0.5 ng/ml but \u0026lt;2 ng/ml indicated moderate risk, \u0026ge;2 ng/ml but \u0026lt; 10 ng/ml indicated high risk, and \u0026ge; 10 ng/ml indicated severe sepsis or septic shock [6-8]. C-reactive protein (CRP) levels were analyzed using the particle-enhanced immunoturbidimetric assay on the Roche Cobas c-311 analyzer, with a cutoff value of CRP \u0026lt;10 mg/L.\u003c/p\u003e\n\u003cp\u003eCerebrospinal fluid (CSF) culture samples were inoculated on chocolate agar, MacConkey agar, and 5% sheep blood agar media, followed by Gram staining. The plates were incubated in an ESCO Cel Culture\u0026reg; CO₂ incubator at 37\u0026deg;C and 5% CO₂ for 24-48 hours. The plates were assessed visually for growth, and Gram staining was performed. The samples were categorized as gram-positive or gram-negative bacteria. Specific microorganism identification and antibiotic susceptibility testing were conducted using the Vitek 2 Compact (BioM\u0026eacute;rieux) system\u003c/p\u003e\n\u003cp\u003eCSF (200 \u0026micro;L) and the hydration solution were added to a Biofire\u0026reg; FilmArray\u0026reg; ME Panel pouch. The prepared pouch was subsequently inserted into the Biofire\u0026reg; FilmArray\u0026reg; PCR automated machine for a one-hour processing cycle. The system automatically extracts and purifies nucleic acids from the lysed sample. Multiplex real-time PCR was conducted in two stages. The first stage involves a high-capacity, massively multiplexed reaction (PCR1), followed by multiple second-stage PCRs (PCR2), which amplify specific sequences from the PCR1 products. The Biofire\u0026reg; FilmArray\u0026reg; real-time PCR system utilizes endpoint-melting curve analysis to identify and generate results for each of the 14 target pathogens represented in the Biofire ME Panel array. The results were automatically interpreted by Biofire\u0026reg; Film Array\u0026reg;\u0026nbsp;software.\u0026nbsp;All procedures were performed in strict accordance with the manufacturer\u0026apos;s instructions to ensure accuracy and reliability.\u003c/p\u003e\n\u003cp\u003eAll the collected data were analyzed using SPSS version 19. The results are expressed as the mean \u0026plusmn; standard deviation, median, and interquartile range for variables such as CRP and PCT. Frequencies and percentages were computed for categorical variables, including sex, CSF culture results, and Biofire ME Panel outcomes. The performance of the Biofire ME Panel was evaluated in terms of sensitivity, specificity, and diagnostic accuracy.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eA total of 132 ICU patients were included in the study, and cerebrospinal fluid (CSF) and blood samples were collected for analysis. Among these patients, 58 (43.9%) were female, and 74 (56.1%) were male. The median age of the patients in the cohort was 38.5 years (IQR: 23\u0026ndash;55). The median C-reactive protein (CRP) level was 3.7 mg/L (IQR: 1.85\u0026ndash;11.75), and the median procalcitonin (PCT) level was 0.10 ng/mL (IQR: 0.025\u0026ndash;0.40).\u003c/p\u003e \u003cp\u003eOf the 132 patients, 9 (6.8%) had positive cultures for \u003cem\u003eListeria monocytogenes\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;6), \u003cem\u003eStreptococcus pneumoniae\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;2), and \u003cem\u003eNeisseria meningitidis\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;1), while 123 patients were culture negative. All 9 culture-positive cases were detected by the Biofire ME Panel, and 114 out of 123 (92.7%) culture-negative cases were confirmed to be true negatives by the Biofire ME Panel. However, among the 10 false-positive patients, 7 (70%) were incorrectly identified as Streptococcus.\u003c/p\u003e \u003cp\u003eThe average time to detect organisms using the Biofire ME Panel assay was 3\u0026thinsp;\u0026plusmn;\u0026thinsp;1 hour after sample collection, whereas CSF culture provided specific microorganism identification and antibiotic susceptibility results within 48\u0026thinsp;\u0026plusmn;\u0026thinsp;2 hours.\u003c/p\u003e \u003cp\u003eIn patients with positive CSF cultures, the mean CRP and PCT levels were significantly elevated, at 61.23\u0026thinsp;\u0026plusmn;\u0026thinsp;20.38 mg/L and 34.47\u0026thinsp;\u0026plusmn;\u0026thinsp;22.78 ng/mL, respectively. In contrast, patients with negative CSF cultures had markedly lower mean C-reactive protein (CRP) and thrombocytocrit (PCT) levels, recorded at 4.73\u0026thinsp;\u0026plusmn;\u0026thinsp;4.27 mg/L and 0.41\u0026thinsp;\u0026plusmn;\u0026thinsp;1.90 ng/mL, respectively.\u003c/p\u003e \u003cp\u003eThe Biofire ME Panel demonstrated strong performance in diagnosing meningitis, with a sensitivity of 100% (95% CI: 63.06\u0026ndash;100%), a specificity of 91.94% (95% CI: 85.67\u0026ndash;96.06%), and an overall diagnostic accuracy of 92.42% (95% CI: 86.51\u0026ndash;96.31%), as shown in Table numbe 1.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDiagnostic accuracy of the Biofire ME Real-Time PCR Assay Compared to Culture\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003en\u0026thinsp;=\u0026thinsp;132\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eCSF Culture\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePositive\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDetected\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8(TP)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10(FP)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNot Detected\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0(FN)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e114(TN)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e124\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBiofire ME Real Time PCR Assay\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSensitivity\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8/8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSpecificity\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e114/124\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e91.94%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDiagnostic accuracy\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8\u0026thinsp;+\u0026thinsp;114/132\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e92.42%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"3\"\u003e* Sensitivity\u0026thinsp;=\u0026thinsp;TP/(TP\u0026thinsp;+\u0026thinsp;FN); specificity\u0026thinsp;=\u0026thinsp;TN/(TN\u0026thinsp;+\u0026thinsp;FP); diagnostic accuracy=(TP\u0026thinsp;+\u0026thinsp;TN)/(TP\u0026thinsp;+\u0026thinsp;TN\u0026thinsp;+\u0026thinsp;FP\u0026thinsp;+\u0026thinsp;FN)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eInfectious meningitis is a serious condition characterized by signs and symptoms such as high-grade fever, nausea, altered consciousness and neck stiffness. These signs can be nonspecific and may overlap with symptoms of other illnesses. While viral meningitis is also common, it is most rapidly fatal (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). Therefore, rapid diagnosis is essential for proper treatment and patient survival. Conventional diagnosis requires laboratory tests such as CSF detailed reports, CSF Gram staining and other biomarkers (PCT, CRP, etc.). However, biomarkers lack specificity, while CSF cultures take several days to obtain results. Recent advancements in the field of molecular diagnostics, especially PCR, are revolutionizing infectious disease management in acute-care settings with rapid, accurate, and potentially cost-effective tools for pathogen detection and antimicrobial resistance profiling (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e)(reference Ammar review). For the diagnosis of meningitis, the rapid diagnostic test Biofire\u0026reg; FilmArray\u0026reg; ME Panel has the capacity to detect 14 different pathogen panels simultaneously based on multiplexed/meclting curve analysis real-time PCR technology and provides rapid results (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThere was a slight male predominance in the cohort studied, with 56.1% males. The median age was 38.50 years, which provided insight into the age distribution of the individuals studied. The median C-reactive protein level was 3.7 mg/L, and the median procalcitonin level was 0.100 ng/mL. In patients in whom CSF culture was positive, the mean CRP and PCT values were substantially greater, indicating that these biomarkers are elevated in patients with confirmed meningitis and suggesting their potential utility in the diagnosis of this disease. This finding is consistent with the expectation that patients with active infections (e.g., meningitis) would have elevated inflammatory marker levels. Similar findings have been reported elsewhere (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe study showed that 8 (6.06%) patients exhibited positive CSF culture growth, indicating bacterial meningitis. \u003cem\u003eListeria monocytogenes\u003c/em\u003e was the most commonly isolated organism, whereas previous studies have identified \u003cem\u003eStreptococcus pneumoniae\u003c/em\u003e as the most common pathogen [12\u0026ndash;13], likely due to differences in climate, hygiene, and population density. The BioFire\u0026reg; real-time PCR ME Panel detected all 8 culture-positive cases, with a sensitivity of 100%. This exceptional sensitivity highlights the panel\u0026rsquo;s efficacy in accurately identifying true-positive cases, thereby ensuring that no cases of bacterial meningitis were missed. Similar findings of high sensitivity were reported by Piccirilli et al. and Lee et al. in their respective studies (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe BioFire\u0026reg; ME Panel demonstrated a specificity of 91.94%, accurately identifying 114 true negatives out of 124 culture-negative specimens, consistent with findings from other studies (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e)(\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e), although Ra`dmard et al. reported a lower specificity of 85.7% (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e)[17]. Importantly, no false negatives were observed, highlighting the strength of the BioFire\u0026reg; ME panel in detecting true-positive cases. However, 10 false positives were identified, indicating that the BioFire\u0026reg; ME panel assay may occasionally detect pathogens where none exist. This suggests the need for confirmatory tests or additional clinical evaluation to prevent unnecessary treatment or invasive procedures, a concern similarly noted by Hansen et al. regarding false positives in their study (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e). Despite its high sensitivity and specificity, clinicians should be cautious of the potential harm that false positives can cause, including unnecessary or potentially toxic treatments.\u003c/p\u003e \u003cp\u003eThis panel screening assay achieved an overall diagnostic accuracy of 92.42%, which combined both sensitivity and specificity. Therefore, this level of accuracy indicates that the panel may have an important role in the diagnosis and management of meningitis within appropriate time limits. CSF culture is still considered the gold standard for diagnosing meningitis; however, this method is typically slow and may miss some bacteria. In contrast, the BioFire\u0026reg; ME Panel provides not only excellent sensitivity and specificity but also a quick turnaround time, providing significant benefit in clinical settings by allowing timely and precise diagnosis.\u003c/p\u003e \u003cp\u003eHowever, despite its excellent performance, the Biofire\u0026reg; ME screening panel has certain limitations. A false positive rate (10/132) could lead to unnecessary treatments and follow-up tests. Furthermore, while CSF culture is regarded as the gold standard, it may not detect all pathogens; therefore, the true performance of the Biofire ME Panel screening assay could vary depending on the actual pathogen distribution across patient groups.\u003c/p\u003e \u003cp\u003eOur study has several limitations. When cerebrospinal fluid (CSF) culture is used as the gold standard, culture methods can yield false negatives, potentially impacting the perceived accuracy of the BioFire\u0026reg; ME Panel. Although the results are promising, further research with larger sample sizes and more diverse populations is necessary to validate the panel\u0026rsquo;s performance across different geographic regions and patient demographics. Additionally, the limited number of samples may have contributed to the absence of certain organisms in the study. Furthermore, this study did not assess how the BioFire\u0026reg; ME Panel results correlated with clinical findings, leaving an important aspect of diagnostic utility unexplored.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn conclusion, our study suggested that the Biofire ME Panel is a valuable tool for diagnosing meningitis, given its high sensitivity, specificity, and overall diagnostic accuracy. This rapid and comprehensive diagnostic method could facilitate prompt and appropriate treatment, ultimately improving patient outcomes.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors have read and approved the manuscript.\u003c/p\u003e\n\u003cp\u003eSD: wrote the main text of manuscript and collect data\u003c/p\u003e\n\u003cp\u003eSN: wrote manuscript and analyze data\u003c/p\u003e\n\u003cp\u003eNA: wrote manuscript and analyze data\u003c/p\u003e\n\u003cp\u003eMAA:critically reviewed the manuscript for intellectual content.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll relevant data are included in the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe approval of Institutional Review Board (IRB)/Ethical Committee of NMC Hospital was taken for study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWritten informed consent was obtained from the all patients.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDedication:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo my Mother (Late Zarina Ramzan Ali), Father (Late Ramzan Ali) and Brother (Late Rehman Ali).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eACKNOWLEDGEMENTS\u003c/strong\u003e\u003cu\u003e:\u003c/u\u003e\u003c/p\u003e\n\u003cp\u003eNone.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eTrujillo-G\u0026oacute;mez J, Tsokani S, Arango-Ferreira C, Atehort\u0026uacute;a-Mu\u0026ntilde;oz S, Jimenez-Villegas MJ, Serrano-Tabares C, et al. Biofire FilmArray Meningitis/Encephalitis panel for the aetiological diagnosis of central nervous system infections: A systematic review and diagnostic test accuracy meta-analysis. EClinicalMedicine. 2022;44:101275.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrouwer MC, Tunkel AR, van de Beek D. Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis. Clin Microbiol Rev. 2010;23(3):467\u0026ndash;92.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLeber AL, Everhart K, Balada-Llasat JM, Cullison J, Daly J, Holt S, et al. Multicenter Evaluation of BioFire FilmArray Meningitis/Encephalitis Panel for Detection of Bacteria, Viruses, and Yeast in Cerebrospinal Fluid Specimens. J Clin Microbiol. 2016;54(9):2251\u0026ndash;61.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAthar MA, Xu Y, Xie X, Xu Z, Ahmad V, Hayder Z, et al. Rapid detection of HCV genotyping 1a, 1b, 2a, 3a, 3b and 6a in a single reaction using two-melting temperature codes by a real-time PCR-based assay. J Virol Methods. 2015;222:85\u0026ndash;90.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamanan P, Bryson AL, Binnicker MJ, Pritt BS, Patel R. Syndromic Panel-Based Testing in Clinical Microbiology. Clin Microbiol Rev. 2018;31(1).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHanson KE. The First Fully Automated Molecular Diagnostic Panel for Meningitis and Encephalitis: How Well Does It Perform, and When Should It Be Used? J Clin Microbiol. 2016;54(9):2222\u0026ndash;4.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTansarli GS, Chapin KC. Diagnostic test accuracy of the BioFire\u0026reg; FilmArray\u0026reg; meningitis/encephalitis panel: a systematic review and meta-analysis. Clin Microbiol Infect. 2020;26(3):281\u0026ndash;90.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHe T, Kaplan S, Kamboj M, Tang YW. Laboratory Diagnosis of Central Nervous System Infection. Curr Infect Dis Rep. 2016;18(11):35.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYang S, Rothman RE. PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings. Lancet Infect Dis. 2004;4(6):337\u0026ndash;48.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUllah MAAVAIUS. Molecular Diagnosis of Hepatitis C Viruses; Technologies and Their Clinical Applications. Microbiol Immunological Commun. 2022;1(1):55\u0026ndash;73.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSager R, Kutz A, Mueller B, Schuetz P. Procalcitonin-guided diagnosis and antibiotic stewardship revisited. BMC Med. 2017;15(1):15.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli SA, Taj MK, Ali SH. Antimicrobial Resistance Pattern of Bacterial Meningitis Among Patients in Quetta, Pakistan. Infect Drug Resist. 2021;14:5107\u0026ndash;20.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee SH, Chen SY, Chien JY, Lee TF, Chen JM, Hsueh PR. Usefulness of the FilmArray meningitis/encephalitis (M/E) panel for the diagnosis of infectious meningitis and encephalitis in Taiwan. J Microbiol Immunol Infect. 2019;52(5):760\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHanson KE, Slechta ES, Killpack JA, Heyrend C, Lunt T, Daly JA, et al. Preclinical Assessment of a Fully Automated Multiplex PCR Panel for Detection of Central Nervous System Pathogens. J Clin Microbiol. 2016;54(3):785\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMessacar K, Breazeale G, Robinson CC, Dominguez SR. Potential clinical impact of the film array meningitis encephalitis panel in children with suspected central nervous system infections. Diagn Microbiol Infect Dis. 2016;86(1):118\u0026ndash;20.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Biofire FilmArray ME Panel, Real-Time PCR, Bacterial Meningitis, CSF Culture","lastPublishedDoi":"10.21203/rs.3.rs-5184548/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5184548/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eMeningitis and encephalitis are critical, life-threatening conditions associated with high rates of morbidity and mortality. Early diagnosis and immediate treatment are essential for decreasing mortality and improving patient outcomes. The Biofire Film Array Meningitis/Encephalitis (ME) panel is a multiplex polymerase chain reaction (PCR)-nucleic acid-based diagnostic assay designed for the rapid diagnosis of bacterial, viral, and fungal pathogens in cerebrospinal fluid. In this study, we evaluated the performance of Biofire Film Array Meningitis/Encephalitis multiplexing real a biofire film array meningitis/encephalitis multiplex real-time PCR assay for the detection of bacterial pathogens in suspected cases of bacterial meningitis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e Cerebrospinal fluid (CSF) and blood samples were collected from 132 intensive care unit (ICU) patients with suspected bacterial meningitis. The clinical samples were analyzed using traditional culture and sensitivity methods, the Biofire Film Array ME Panel multiplex PCR, procalcitonin (PCT), and C-reactive protein (CRP) assays. The sensitivity, specificity and diagnostic accuracy of the Biofire Film Array ME Panel real-time PCR assay were evaluated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eA total of 132 patients, with a mean age of 38.9 ± 23.7 years, were included in the study. The mean levels of C-reactive protein (CRP) and procalcitonin (PCT) were 52.41 ± 82.23 mg/L and 3.7 ± 7.6 ng/mL, respectively. Among these patients, 9 (6.8%) had positive cultures for \u003cem\u003eListeria monocytogenes\u003c/em\u003e (n = 6), \u003cem\u003eStreptococcus pneumonia\u003c/em\u003e (n = 2), and \u003cem\u003eNeisseria meningitides\u003c/em\u003e (n = 1), while 123 patients were culture negative.\u003c/p\u003e\n\u003cp\u003eAll 9 culture-positive cases were detected by the Biofire ME Panel, and 114 out of 123 (92.7%) culture-negative cases were confirmed as true negatives by the panel. The Biofire ME Panel demonstrated excellent diagnostic performance, with a sensitivity of 100% (95% CI: 63.06–100%), a specificity of 91.94% (95% CI: 85.67–96.06%), and an overall diagnostic accuracy of 92.42% (95% CI: 86.51–96.31%).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: The Biofire ME Panel is strongly correlatedwith CSF culture and has excellent diagnostic accuracy for bacterial meningitis. Moreover, the NRS-2002 may be used as a routine test for suspected cases of bacterial meningitis for early diagnosis and optimal treatment to reduce mortality and morbidity.\u003c/p\u003e","manuscriptTitle":"Evaluation of the Performance of a Biofire FilmArray Meningitis/Encephalitis Multiplex Real-Time PCR Assay for Bacterial Meningitis Diagnosis at a Tertiary Care Hospital","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-08 09:17:42","doi":"10.21203/rs.3.rs-5184548/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"119189a9-c622-49c3-99da-d1fa42f893eb","owner":[],"postedDate":"October 8th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-10-19T06:53:10+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-08 09:17:42","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5184548","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5184548","identity":"rs-5184548","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}