Journal of Clinical Virology最新文献

{"title":"Does the ultrasensitive HBsAg Next assay enhance Hepatitis B diagnosis? An evaluation of analytical performances","authors":"Joachim Bourdin ,&nbsp;Pierre Sellier ,&nbsp;Maud Salmona ,&nbsp;Caroline Lascoux-Combe ,&nbsp;Constance Delaugerre ,&nbsp;Sarah Maylin","doi":"10.1016/j.jcv.2024.105707","DOIUrl":"10.1016/j.jcv.2024.105707","url":null,"abstract":"<div><h3>Background</h3><p>Accurate laboratory confirmation for Hepatitis B diagnosis and monitoring are crucial. Recently an ultrasensitive immunoassay test, the HBsAg Next (HBsAgNx), has been reported approximately eight times more sensitive than current HBsAg assays. The aim of our study was to assess the analytical performances of this new test.</p></div><div><h3>Methodology</h3><p>253 clinical samples from Saint Louis University Hospital were analyzed, splitted into four panels: (<span>1</span>) routine prospectively screening serums (<em>n</em> = 196), (<span>2</span>) retrospective serum samples before HBV reactivation (HBV-R) (<em>n</em> = 18), (<span>3</span>) occult HBV infection (OBI) (<em>n</em> = 10) and (<span>4</span>) a selection of wild type HBV genotypes (<em>n</em> = 29)</p></div><div><h3>Results</h3><p>Panel 1, showed robust agreement with the HBsAg Qualitative II (HBsAgQII) assay (Cohen's kappa = 0.83). Despite this agreement, 7 false positive with the HBsAgQII assay were found negative with HBsAgNx. One OBI was detected only with HBsAgNx. Panel 2 showed potential time savings in diagnosing HBV-R using HBsAgNx among 4/18 HBsAg positives samples. Panel 3 highlighted the ability of HBsAgNx to detect HBsAg in OBI patients defined by negative for HBsAg with HBsAgQII assay and positive for HBV DNA. Furthermore, the HBsAgNx assay detected all different genotypes.</p></div><div><h3>Conclusion</h3><p>The study highlights the effectiveness of the HBsAgNx assay, showing its performance. It excels in detecting weakly positive samples and addressing challenging cases. HBsAgNx assay demonstrates promising analytical performances, with improved sensitivity and specificity compared to standard HBsAgQII assay, able to detect all genotypes. Its potential impact on early detecting and monitoring reactivations, and occult infections could be very useful in clinical practice.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"174 ","pages":"Article 105707"},"PeriodicalIF":4.0,"publicationDate":"2024-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141534561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Performance of a rapid recency assay for detection of early HIV infection","authors":"Clara Di Germanio ,&nbsp;Xutao Deng ,&nbsp;Eduard Grebe ,&nbsp;Jeffrey A. Johnson ,&nbsp;Silvina Masciotra ,&nbsp;Michael P. Busch ,&nbsp;Philip J. Norris","doi":"10.1016/j.jcv.2024.105708","DOIUrl":"10.1016/j.jcv.2024.105708","url":null,"abstract":"<div><p>The Asanté HIV-1 Rapid Recency assay's ‘verification’ line detected HIV infection a median of 18 days later than a nucleic acid detection assay and performed similarly to 19 other existing rapid HIV antibody tests. Pending regulatory approval, the assay could be an option with other rapid tests in national HIV-1 testing algorithms, which would allow collection of HIV recency data as part of a national screening program without requiring additional testing.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"174 ","pages":"Article 105708"},"PeriodicalIF":4.0,"publicationDate":"2024-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141468450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Direct whole-genome sequencing of HIV-1 for clinical drug-resistance analysis and public health surveillance","authors":"Rosa C. Coldbeck-Shackley ,&nbsp;Penelope J. Adamson ,&nbsp;Daryn Whybrow ,&nbsp;Caitlin A. Selway ,&nbsp;Lito E. Papanicolas ,&nbsp;Mark Turra ,&nbsp;Lex E.X. Leong","doi":"10.1016/j.jcv.2024.105709","DOIUrl":"10.1016/j.jcv.2024.105709","url":null,"abstract":"<div><h3>Background</h3><p>Human Immunodeficiency virus type 1 (HIV-1) remains a significant global health threat partly due to its ability to develop resistance to anti-retroviral therapies. HIV-1 genotype and drug resistance analysis of the polymerase (<em>pol)</em> sequence is a mainstay of its clinical and public health management. However, as new treatments and resistances evolve, analysis methods must change accordingly. In this study, we outline the development and implementation of a direct whole-genome sequencing approach (dWGS) using probe-capture target-enrichment for HIV-1 genotype and drug resistance analysis.</p></div><div><h3>Methods</h3><p>We implemented dWGS and performed parallel <em>pol</em> Sanger sequencing for clinical samples, followed by comparative genotype and drug-resistance analysis. These HIV-1 WGS sequences were also utilised for a novel partitioned phylogenetic analysis.</p></div><div><h3>Results</h3><p>Optimised nucleic acid extraction and DNAse I treatment significantly increased HIV-1 whole-genome coverage and depth, and improved recovery of high-quality genomes from low viral load clinical samples, enabling routine sequencing of viral loads as low as 1000 copies/mL. Overall, dWGS was robust, accurate and more sensitive for detecting low-frequency variants at drug-resistance sites compared to Sanger sequencing. Analysis of multiple sequence regions improved phylogenetic reconstruction for recombinant HIV-1 sequences compared to analysis of <em>pol</em> sequence alone.</p></div><div><h3>Conclusions</h3><p>These findings demonstrate dWGS enhances HIV-1 drug-resistance analysis by quantitative variant detection and improves reconstruction of HIV-1 phylogenies compared to traditional <em>pol</em> sequencing. This work supports that HIV-1 dWGS is a viable option to replace Sanger sequencing for clinical and public health applications.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"174 ","pages":"Article 105709"},"PeriodicalIF":4.0,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1386653224000714/pdfft?md5=7ed4ba8f5f74ab1a945c420c41a73fc3&pid=1-s2.0-S1386653224000714-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinical performance of the Roche Cobas Liat SARS-CoV-2 & influenza A/B assay: A systematic review and meta-analysis","authors":"Eunjin Chang ,&nbsp;Kibum Jeon ,&nbsp;Nuri Lee ,&nbsp;Min-Jeong Park ,&nbsp;Wonkeun Song ,&nbsp;Hyun Soo Kim ,&nbsp;Han-Sung Kim ,&nbsp;Jae-Seok Kim ,&nbsp;Jimin Kim ,&nbsp;Seri Jeong","doi":"10.1016/j.jcv.2024.105706","DOIUrl":"10.1016/j.jcv.2024.105706","url":null,"abstract":"<div><p>Respiratory tract infections caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza viruses are persistent and critical. The Cobas Liat SARS-CoV-2 &amp; influenza A/B assay (Multiplex Liat), the FDA-authorized point-of-care reverse transcriptase polymerase chain reaction (RT-PCR) assay, has a turnaround time of 20 min and high accuracy. This study evaluates the pooled performance of this assay to provide practical information. This meta-analysis was registered in PROSPERO (registration number: CRD42023467579). A systematic literature search was conducted within PubMed, Ovid-EMBASE, and the Cochrane Library for articles evaluating the accuracy of the Multiplex Liat assay through September 2023. A random-effects model was used to calculate the pooled diagnostic values with real-time RT-PCR (rRT-PCR) as a reference test. A total of 4,705 samples from eight studies were included in the primary meta-analysis. The overall pooled sensitivity and specificity of Multiplex Liat were 100.0 % (95 % confidence interval [CI] = 96.7 %–100.0 %) and 99.7 % (95 % CI = 98.7 %–99.9 %), respectively. The presence of variants of concern or in-house rRT-PCR assays as reference standards did not significantly affect the pooled diagnostic performance of the Multiplex Liat. When 5,333 samples from nine studies were assessed for sensitivity, the pooled sensitivity was 100.0 % (95 % CI = 85.8 %–100.0 %) without a significant difference. This meta-analysis demonstrates the usefulness of Multiplex Liat for the detection of SARS-CoV-2 based on pooled diagnostic values. These practical findings may facilitate appropriate settings for the diagnosis and management of patients with respiratory tract infections.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"174 ","pages":"Article 105706"},"PeriodicalIF":4.0,"publicationDate":"2024-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1386653224000684/pdfft?md5=3155d214245d6a8c7954b3981595fcb1&pid=1-s2.0-S1386653224000684-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141389457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Melting curve analysis reveals false-positive norovirus detection in a molecular syndromic panel","authors":"Nancy Matic ,&nbsp;Tanya Lawson ,&nbsp;Matthew Young ,&nbsp;Willson Jang ,&nbsp;Jennifer Bilawka ,&nbsp;Leah Gowland ,&nbsp;Gordon Ritchie ,&nbsp;Victor Leung ,&nbsp;Michael Payne ,&nbsp;Aleksandra Stefanovic ,&nbsp;Marc G. Romney ,&nbsp;Christopher F. Lowe","doi":"10.1016/j.jcv.2024.105697","DOIUrl":"10.1016/j.jcv.2024.105697","url":null,"abstract":"<div><h3>Background</h3><p>Molecular syndromic panels can improve rapidity of results and ease clinical laboratory workflow, although caution has been raised for potential false-positive results. Upon implementation of a new panel for infectious diarrhea (BioFire® FilmArray® Gastrointestinal [GI] Panel, bioMérieux) in our clinical laboratory, a higher than expected number of stool samples with norovirus were detected.</p></div><div><h3>Objectives</h3><p>The goal of this study was to investigate positive percent agreement and the false-positive rate of norovirus detected by the multiplex BioFire GI panel compared to a singleplex commercial assay.</p></div><div><h3>Study design</h3><p>From October 2023 to January 2024, all prospective stool samples with a positive norovirus result by BioFire had melting curves reviewed manually using the BioFire FilmArray Torch System. Stool samples further underwent testing by a supplementary real-time RT-PCR assay (Xpert® Norovirus, Cepheid) for comparative analysis.</p></div><div><h3>Results</h3><p>Of the 50 stool samples with norovirus detected by BioFire, 18 (36 %) tested negative by Xpert (deemed \"false-positives\"). Furthermore, melting curve analysis revealed nearly all of these samples had atypical melting curve morphologies for the \"Noro-1\" target on BioFire (16/18, 89 %), which was statistically significant (Odds Ratio 173.2, 95 % CI [22.2, 5326.9], <em>p</em> &lt; 0.0001). Stool samples with multiple pathogens detected by BioFire including norovirus were not more likely to produce false-positive norovirus results (Odds Ratio 1, 95 % CI [0.3, 3.3], <em>p</em> = 1).</p></div><div><h3>Conclusions</h3><p>Although not described in the manufacturer's <em>Instructions for Use</em>, we propose routine manual review of melting curves for the BioFire GI panel prior to reporting, to mitigate potential false-positive norovirus results.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"173 ","pages":"Article 105697"},"PeriodicalIF":8.8,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1386653224000593/pdfft?md5=c423b5c837627ec86baa64edd7c77889&pid=1-s2.0-S1386653224000593-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141183853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multicenter benchmarking of short and long read wet lab protocols for clinical viral metagenomics","authors":"F. Xavier Lopez-Labrador ,&nbsp;Michael Huber ,&nbsp;Igor A. Sidorov ,&nbsp;Julianne R. Brown ,&nbsp;Lize Cuypers ,&nbsp;Lies Laenen ,&nbsp;Bert Vanmechelen ,&nbsp;Piet Maes ,&nbsp;Nicole Fischer ,&nbsp;Ian Pichler ,&nbsp;Nathaniel Storey ,&nbsp;Laura Atkinson ,&nbsp;Stefan Schmutz ,&nbsp;Verena Kufner ,&nbsp;Sander van Boheemen ,&nbsp;Claudia E. Mulders ,&nbsp;Adam Grundhoff ,&nbsp;Patrick Blümke ,&nbsp;Alexis Robitaille ,&nbsp;Ondrej Cinek ,&nbsp;Jutte J.C. de Vries","doi":"10.1016/j.jcv.2024.105695","DOIUrl":"10.1016/j.jcv.2024.105695","url":null,"abstract":"<div><p>Metagenomics is gradually being implemented for diagnosing infectious diseases. However, in-depth protocol comparisons for viral detection have been limited to individual sets of experimental workflows and laboratories. In this study, we present a benchmark of metagenomics protocols used in clinical diagnostic laboratories initiated by the European Society for Clinical Virology (ESCV) Network on NGS (ENNGS).</p><p>A mock viral reference panel was designed to mimic low biomass clinical specimens. The panel was used to assess the performance of twelve metagenomic wet lab protocols currently in use in the diagnostic laboratories of participating ENNGS member institutions. Both Illumina and Nanopore, shotgun and targeted capture probe protocols were included. Performance metrics sensitivity, specificity, and quantitative potential were assessed using a central bioinformatics pipeline.</p><p>Overall, viral pathogens with loads down to 10⁴ copies/ml (corresponding to C_T values of 31 in our PCR assays) were detected by all the evaluated metagenomic wet lab protocols. In contrast, lower abundant mixed viruses of C_T values of 35 and higher were detected only by a minority of the protocols. Considering the reference panel as the gold standard, optimal thresholds to define a positive result were determined per protocol, based on the horizontal genome coverage. Implementing these thresholds, sensitivity and specificity of the protocols ranged from 67 to 100 % and 87 to 100 %, respectively.</p><p>A variety of metagenomic protocols are currently in use in clinical diagnostic laboratories. Detection of low abundant viral pathogens and mixed infections remains a challenge, implying the need for standardization of metagenomic analysis for use in clinical settings.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"173 ","pages":"Article 105695"},"PeriodicalIF":8.8,"publicationDate":"2024-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S138665322400057X/pdfft?md5=22ce2b90dc93fb6b9779951463075d74&pid=1-s2.0-S138665322400057X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141186679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Shedding of measles vaccine RNA in children after receiving measles, mumps and rubella vaccination","authors":"Matthew C. Washam ,&nbsp;Amy L. Leber ,&nbsp;Sophonie J. Oyeniran ,&nbsp;Kathy Everhart ,&nbsp;Huanyu Wang","doi":"10.1016/j.jcv.2024.105696","DOIUrl":"10.1016/j.jcv.2024.105696","url":null,"abstract":"<div><h3>Background</h3><p>Measles, mumps, and rubella(MMR) vaccination is critical to measles outbreak responses. However, vaccine reactions and detection of measles vaccine RNA in recently immunized persons may complicate case classification especially in those presenting with another respiratory viral illness. We aim to characterize cases of measles vaccine shedding in recently vaccinated children presenting with respiratory viral symptoms.</p></div><div><h3>Methods</h3><p>Children who were tested with a multiplex respiratory panel &lt;30 days after receiving MMR were identified. Remnant nasopharyngeal(NP) samples were tested for measles vaccine by PCR. Medical records were reviewed for demographics, presenting symptoms, and test results.</p></div><div><h3>Results</h3><p>From January 2022 to March 2023, 127 NP from children who received MMR were tested. Ninety-six NP were collected after the first dose, of which 33(34.4 %) were positive for vaccine RNA. The median interval between MMR and detection was 11.0 days. Thirty-one NP were collected after the second MMR and 1(3.2 %) was positive; time between the vaccination and detection was 18.9 days. Median cycle threshold(Ct) value of the measles PCR for vaccine shedding was significantly higher than median Ct in children with wild-type infection.</p></div><div><h3>Conclusion</h3><p>Shedding of measles vaccine RNA is not uncommon and vaccine RNA can be detected up to 29 days post MMR; the amount of vaccine RNA shedding is low indicated by high Ct values. Clinicians and public health officials should consider performing measles vaccine testing on those testing positive for measles within one month of MMR vaccination, especially if the Ct value is high and definitive epidemiological links are absent.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"173 ","pages":"Article 105696"},"PeriodicalIF":8.8,"publicationDate":"2024-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141145599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Resurgence of common respiratory viruses in patients with community-acquired pneumonia (CAP)—A prospective multicenter study","authors":"Theo Dähne ,&nbsp;Wolfgang Bauer ,&nbsp;Andreas Essig ,&nbsp;Bernhard Schaaf ,&nbsp;Grit Barten-Neiner ,&nbsp;Christoph D. Spinner ,&nbsp;Mathias W. Pletz ,&nbsp;Gernot Rohde ,&nbsp;Jan Rupp ,&nbsp;Martin Witzenrath ,&nbsp;Marcus Panning","doi":"10.1016/j.jcv.2024.105694","DOIUrl":"https://doi.org/10.1016/j.jcv.2024.105694","url":null,"abstract":"<div><h3>Background</h3><p>Community-acquired pneumonia (CAP) is a major global cause of death and hospitalization. Bacteria or community-acquired viruses (CARVs) cause CAP. COVID-19 associated restrictions effectively reduced the circulation of CARVs.</p></div><div><h3>Objectives</h3><p>The aim of this study was to analyze the proportion of CARVs in adult patients with CAP from mid-2020 to mid-2023. Specifically, we aimed to compare the rate of influenza virus, SARS-CoV-2, and RSV detections in patients aged 18–59 years and ≥60 years.</p></div><div><h3>Study design</h3><p>We analyze the proportion of 21 community-acquired respiratory viruses (CARVs) and three atypical bacteria (<em>Bordetella pertussis, Legionella pneumophila</em>, and <em>Mycoplasma pneumoniae</em>) in nasopharyngeal swab samples using molecular multiplex methods within the prospective, multicentre, multinational study of the German study Group CAPNETZ. We used stringent inclusion criteria throughout the study.</p></div><div><h3>Results</h3><p>We identified CARVs in 364/1,388 (26.2 %) patients. In detail, we detected SARS-CoV-2 in 210/1,388 (15.1 %), rhino-/enterovirus in 64/1,388 (4.6 %), influenza virus in 23/1,388 (1.6 %) and RSV in 17/1,388 (1.2 %) of all patients. We detected RSV and influenza more frequently in patients ≥60 years, especially in 22/23 compared to the previous season. None of the atypical bacteria were detected.</p></div><div><h3>Conclusions</h3><p>Beginning in 2023, we demonstrate a re-emergence of CARVs in CAP patients. Effective vaccines or specific antiviral therapies for more than two thirds of the detected viral infections are currently available. High detection rates of vaccine-preventable viruses in older age groups support targeted vaccination campaigns.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"173 ","pages":"Article 105694"},"PeriodicalIF":8.8,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1386653224000568/pdfft?md5=2f2377eb7ebe1dce67f90f74cb282d26&pid=1-s2.0-S1386653224000568-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141083391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analytical and clinical validation of a novel, laboratory-developed, modular multiplex-PCR panel for fully automated high-throughput detection of 16 respiratory viruses","authors":"Hui Ting Tang ,&nbsp;Dominik Nörz ,&nbsp;Moritz Grunwald,&nbsp;Katja Giersch,&nbsp;Susanne Pfefferle,&nbsp;Nicole Fischer,&nbsp;Martin Aepfelbacher,&nbsp;Holger Rohde,&nbsp;Marc Lütgehetmann","doi":"10.1016/j.jcv.2024.105693","DOIUrl":"10.1016/j.jcv.2024.105693","url":null,"abstract":"<div><h3>Background</h3><p><strong>:</strong> Viral respiratory Infections pose a health risk, especially to vulnerable patient populations. Effective testing programs can detect and differentiate these infections at an early stage, which is particularly important for high-risk clinical departments. The objective of this study was to develop and validate a multiplex PCR-panel for 16 different respiratory viruses on a fully-automated high-throughput platform.</p></div><div><h3>Methods</h3><p>Three multiplex-PCR assays were designed to run on the cobas5800/6800/8800 systems, consolidating 16 viral targets: RESP1: SARS-CoV-2, influenza-A/B, RSV; RESP2: hMPV, hBoV, hAdV, rhino-/ENV; RESP3: HPIV-1–4, hCoV-229E, hCoV-NL63, hCoV-OC43, hCoV-HKU1. Analytic performance was evaluated using digital-PCR based standards and international reference material. Clinical performance was determined by comparing results from clinical samples with reference assays.</p></div><div><h3>Results</h3><p>Analytical sensitivity (i.e. lower limit of detection (LoD), 95 % probability of detection) was determined as follows: SARS-CoV-2: 29.3 IU/ml, influenza-A: 179.9 cp/ml, influenza-B: 333.9 cp/ml and RSV: 283.1 cp/ml. LoDs of other pathogens ranged between 9.4 cp/ml (hCoV-NL63) and 21,419 cp/ml (HPIV-2). Linearity was verified over 4–7 log-steps with pooled standard differentials (SD) ranging between 0.18–0.70ct. Inter-/intra-run variability (precision) was assessed for all targets over 3 days. SDs ranged between 0.13–0.74ct. Positive agreement in clinical samples was 99.4 % and 95 % for SARS-CoV-2 and influenza-A respectively. Other targets were in the 80–100 % range. Negative agreement varied between 96.3–100 %.</p></div><div><h3>Discussion</h3><p>Lab-developed tests are a key factor for effective clinical diagnostics. The multiplex panel presented in this study demonstrated high performance and provides an easily scalable high-throughput solution for respiratory virus testing, e.g. for testing in high-risk patient populations.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"173 ","pages":"Article 105693"},"PeriodicalIF":8.8,"publicationDate":"2024-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1386653224000556/pdfft?md5=6da38b25c92dba819b4157dad492192a&pid=1-s2.0-S1386653224000556-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141039866","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinical presentation, viral shedding, and neutralizing antibody responses of mpox cases in South Korea: Single center experience","authors":"BumSik Chin ,&nbsp;Jihye Um ,&nbsp;Min-Kyung Kim ,&nbsp;Hyang Su Kim ,&nbsp;Hong Soon Yim ,&nbsp;Hyee Jin Cho ,&nbsp;So Yun Lim ,&nbsp;Yeonjae Kim ,&nbsp;Jaehyun Jeon ,&nbsp;Jun-Sun Park","doi":"10.1016/j.jcv.2024.105692","DOIUrl":"10.1016/j.jcv.2024.105692","url":null,"abstract":"<div><h3>Background</h3><p>A global mpox outbreak occurred in 2022, and a domestic outbreak started in South Korea in April 2023. This study aimed to evaluate the clinical characteristics, viral shedding, and immune response of mpox in South Korea.</p></div><div><h3>Methods</h3><p>Patients hospitalized with mpox in the National Medical Center between September 2022 and June 2023 were included in this study. Oropharyngeal (OP), anogenital lesion (AL), and skin lesion (SL) swabs and blood samples were collected, and monkeypox virus (MPXV) DNA using real-time polymerase chain reaction (RT-PCR) and culture assays were performed. Neutralizing antibodies (NAbs) against MPXV A.2.1, B.1.1, and B.1.3 were detected using plaque reduction neutralization tests.</p></div><div><h3>Results</h3><p>Eighteen patients were enrolled, of whom 17 (94.4 %) were male, with a median (IQR) age of 32.5 (24–51) years. While nine (50 %) were HIV-infected individuals, none of them revealed CD4+ counts less than 200 cells/μL. MPXV DNA was detected in 87.3 % and 82.7 % of patient's ALs and SLs, respectively, until 2 weeks after symptom onset. While MPXV was isolated for up to 15 days in all three sample types, the culture positivity decreased to 53.8 % and 42.9 % in ALs and SLs after 10 days, respectively, and 28.6 % and 22.2 %, respectively, after 2 weeks from symptom onset. The NAb titers against MPXV A.2.1 were significantly lower than those against B.1.1 and B.1.3.</p></div><div><h3>Conclusions</h3><p>Infectious MPXV was isolated from various anatomical sites up to 15 days after symptom onset. The MPXV NAb response was varied among different lineages, and this implies limited cross-lineage protection.</p></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"173 ","pages":"Article 105692"},"PeriodicalIF":8.8,"publicationDate":"2024-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1386653224000544/pdfft?md5=72fb19102f3a57fd2831abe390bf7d9a&pid=1-s2.0-S1386653224000544-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141028829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}