Journal of Clinical Virology最新文献_第8页

False-positive results in fourth-generation HIV screening tests: Prevalence and associated factors in Sichuan, a high HIV burden province of China 第四代HIV筛查假阳性结果：中国HIV高负担省份四川的流行情况及相关因素

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-08-01 Epub Date: 2025-07-01 DOI: 10.1016/j.jcv.2025.105831

Hong Zhang , Jiaqiang Wang , Xiangqin Liu , Xueru Li , Xuexi Zeng , Qing Luo , Jialing Zhong

Immunodeficiency virus (HIV) antigen/antibody screening assays are highly sensitive and specific, but false-positive (FP) results remain a challenge. Understanding the prevalence and factors associated with these FP results is crucial, especially in high HIV burden regions. A retrospective cohort study of 370,291 patients screened with the ARCHITECT HIV Ag/Ab Combo assay at a Sichuan tertiary hospital (January 2022–December 2023) was conducted. We calculated HIV prevalence and assessed the test's FP rate, sensitivity, specificity and positive predictive value (PPV). Clinical characteristics and associated disease profiles of individuals with FP results were also analyzed. The overall HIV infection rate was 0.17 %. The FP rate for HIV screening was 0.08 %, with higher incidences observed among females, children (aged 0–17 years), and individuals aged 66 and older (P < 0.001). The mean signal-to-cutoff ratio (S/CO) in true positives (TPs) was significantly higher than that in FPs (576.63 vs. 1.94, P < 0.0001). A receiver operating characteristic (ROC)-determined cutoff of 19.6 provided optimal sensitivity (95.10 %) and specificity (99.99 %). FP results were associated with 18 disease categories, with digestive system disorders being the most prevalent. Malignant tumors, pregnancy, and cerebral infarction were also linked to FPs. These findings highlight the critical need for targeted screening strategies and more precise interpretation protocols to improve diagnostic accuracy. Furthermore, the link between FP results and various non-HIV-related diseases suggests that careful patient characterization may aid in identifying underlying conditions, thereby informing more effective clinical decision-making and public health interventions.

免疫缺陷病毒（HIV）抗原/抗体筛选测定是高度敏感和特异性的，但假阳性（FP）结果仍然是一个挑战。了解这些计划生育结果的流行情况和相关因素至关重要，特别是在艾滋病毒高负担地区。对四川省某三级医院（2022年1月- 2023年12月）采用ARCHITECT HIV Ag/Ab组合检测筛查的370,291例患者进行回顾性队列研究。我们计算HIV流行率，并评估该检测的FP率、敏感性、特异性和阳性预测值（PPV）。分析了FP结果个体的临床特征和相关疾病概况。总体HIV感染率为0.17%。HIV筛查的计划生育率为0.08%，在女性、儿童（0-17岁）和66岁及以上的人群中发病率较高(P <；0.001)。真阳性（TPs）的平均信号截止比（S/CO）显著高于FPs (576.63 vs. 1.94, P <；0.0001)。受试者工作特征（ROC）确定的截止值为19.6，提供了最佳的灵敏度（95.10%）和特异性（99.99%）。FP结果与18种疾病相关，其中消化系统疾病最为普遍。恶性肿瘤、妊娠和脑梗死也与FPs有关。这些发现强调了有针对性的筛查策略和更精确的解释方案的迫切需要，以提高诊断的准确性。此外，计划生育结果与各种非艾滋病毒相关疾病之间的联系表明，仔细描述患者特征可能有助于确定潜在疾病，从而为更有效的临床决策和公共卫生干预提供信息。

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引用次数: 0

Comparative evaluation of STANDARD™ M10 Flu/RSV/SARS-CoV-2 and Savanna® Respiratory Viral Panel-4 assays for the rapid molecular diagnosis of influenza A/B virus, respiratory syncytial virus and SARS-CoV-2 用于流感A/B病毒、呼吸道合胞病毒和SARS-CoV-2快速分子诊断的STANDARD™M10 Flu/RSV/SARS-CoV-2和Savanna®Respiratory Viral Panel-4检测的比较评价

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-08-01 Epub Date: 2025-06-11 DOI: 10.1016/j.jcv.2025.105827

Juulia Suominen, Raisa Loginov, Hannimari Kallio-Kokko

Background

The occurrence of respiratory infections caused by seasonal viruses influenza A/B, RSV and SARS-CoV-2 has increased the demand for rapid diagnostic assays. Comparative performance data of such assays is required.

Methods

In this retrospective study, clinical samples were tested with the STANDARD™ M10 Flu/RSV/SARS-CoV-2 test and the novel Savanna® Respiratory Viral Panel-4 tests, with Xpert® Xpress SARS-CoV-2/Flu/RSV as the reference. All three are RT-PCR tests suitable for point-of-care testing. Discordant results on the Savanna assay were retested with a new research-use-only protocol. Serial dilution testing for all three was performed with an external control.

Results

A total of 141 clinical samples, including 106 specimens positive for at least one virus, were analyzed. The M10 assay showed sensitivities of 100 %, 95.7 %, 97.1 % and 97.0 % for influenza A, B, RSV and SARS-CoV-2, respectively. The Savanna assay showed sensitivities of 92.6 %, 95.7 %, 100 % and 90.9 %. Both assays exhibited high specificity (≥99 %), except for the Savanna assay’s lower specificity for RSV (94.2 %) and SARS-CoV-2 (94.3 %). Savanna had a higher retest rate (5.0 %), while M10 produced only conclusive results. Serial dilution testing showed that Xpert detected three viruses more effectively than the other assays.

Conclusion

Both M10 and Savanna performed well for influenza A/B, but M10 was superior for RSV and SARS-CoV-2 due to false positives with Savanna. The new Savanna protocol showed promise, but further studies are required to confirm these findings. Xpert assay was the most sensitive for detecting low viral amounts.

季节性流感A/B、RSV和SARS-CoV-2引起的呼吸道感染的发生增加了对快速诊断检测的需求。需要这些分析的比较性能数据。方法以Xpert®Xpress SARS-CoV-2/Flu/RSV /RSV为对照，采用STANDARD™M10 Flu/RSV/SARS-CoV-2检测试剂盒和新型Savanna®Respiratory Viral Panel-4检测试剂盒对临床样本进行回顾性检测。这三种方法都是适用于即时检测的RT-PCR方法。用一种新的仅供研究使用的方案重新测试了稀树草原试验中不一致的结果。用外部对照对这三种药物进行连续稀释试验。结果共检出141份临床标本，其中至少一种病毒阳性106份。M10试验对甲型流感、乙型流感、RSV和SARS-CoV-2的敏感性分别为100%、95.7%、97.1%和97.7%。稀树草原试验的灵敏度分别为92.6%、95.7%、100%和90.9%。除了Savanna法对RSV（94.2%）和SARS-CoV-2（94.3%）的特异性较低外，两种检测方法均表现出高特异性（≥99%）。Savanna的复验率较高（5.0%），而M10仅产生结论性结果。系列稀释试验表明，Xpert比其他试验更有效地检测到三种病毒。结论M10和Savanna对流感A/B均有较好的检测效果，而M10对RSV和SARS-CoV-2均有较好的检测效果。新的热带草原方案显示出了希望，但需要进一步的研究来证实这些发现。Xpert法对检测低病毒量最敏感。

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引用次数: 0

Implications of subtyping influenza A amongst H5N1 surveillance efforts 甲型流感亚型在H5N1监测工作中的意义

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-08-01 Epub Date: 2025-05-29 DOI: 10.1016/j.jcv.2025.105810

Samuel M. Goodfellow, Jennifer Dien Bard, Cristina Costales

引用次数: 0

The Human Papillomavirus (HPV) Laboratory e-Manual: A comprehensive guide for HPV testing and research 人类乳头瘤病毒（HPV）实验室电子手册：HPV检测和研究的综合指南

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-08-01 Epub Date: 2025-07-10 DOI: 10.1016/j.jcv.2025.105834

Laila Sara Arroyo Mühr , Carina Eklund , Camilla Lagheden , Rita Mariel Correa , María Dolores Fellner , Maria Alejandra Picconi , Nazlı Songur , Murat Gultekin , Kate Cuschieri , Jean-Luc Prétet , Quentin Lepiller , Alice Baraquin , Steffi Silling , Kristiane Søreng , Milan Stosic , Gro Kummeneje Presthus , Marc Arbyn , Michael Peeters , Steven Van Gucht , Elizaveta Padalko , Joakim Dillner

Background

Human Papillomavirus (HPV) vaccination and HPV-based cervical cancer screening are central pillars of the World Health Organization (WHO) global cervical cancer elimination strategy. The WHO HPV Laboratory Manual, published in 2009, has provided essential guidance to promote an internationally comparable quality of HPV testing for many years. As the development in this area is rapid, the Global Network of National HPV Reference Laboratories considered that there is a need for an updated HPV Laboratory e-Manual to serve as a comprehensive and interactive resource for professionals engaged in quality-assured HPV testing for research and/or HPV-based cancer control.

Content

The HPV Laboratory e-Manual covers key areas, including laboratory quality assurance, HPV taxonomy and risk association, collection and handling of specimens, nucleic acid extraction, HPV detection and typing, HPV serology, data management, and the use of international standards. It provides up-to-date protocols and best practices to enhance accuracy and reliability of HPV testing. Interactive features allow for real-time updates, making it a dynamic resource for laboratories worldwide. The e-Manual is freely available at: https://www.hpvcenter.se/hpv-laboratory-manual/.

Collaborators

The e-Manual has been developed by international experts from 11 countries, including contributors from the International HPV Reference Center (IHRC, Sweden), the CDC’s Global HPV Reference Laboratory (USA), and multiple National HPV Reference Laboratories (NRLs). The standard procedure for writing a chapter was that 2 NRLs authored the chapter and 1 other NRL reviewed it.

Conclusion

The HPV Laboratory e-Manual represents a step toward global harmonization in laboratory methodologies for HPV testing, underpinning both research and cervical cancer control efforts.

人类乳头瘤病毒（HPV）疫苗接种和基于HPV的宫颈癌筛查是世界卫生组织（WHO）全球消除宫颈癌战略的中心支柱。世卫组织2009年出版的《人乳头瘤病毒实验室手册》多年来为促进具有国际可比性的人乳头瘤病毒检测质量提供了基本指导。由于这一领域的发展迅速，国家HPV参考实验室全球网络认为有必要更新HPV实验室电子手册，为从事有质量保证的HPV检测研究和/或基于HPV的癌症控制的专业人员提供全面和互动的资源。HPV实验室电子手册涵盖了关键领域，包括实验室质量保证、HPV分类和风险关联、标本收集和处理、核酸提取、HPV检测和分型、HPV血清学、数据管理和国际标准的使用。它提供了最新的协议和最佳做法，以提高HPV检测的准确性和可靠性。交互功能允许实时更新，使其成为全球实验室的动态资源。电子手册由来自11个国家的国际专家编写，其中包括来自国际HPV参考中心（IHRC，瑞典）、疾病预防控制中心全球HPV参考实验室（美国）和多个国家HPV参考实验室（nrl）的贡献者。写一章的标准程序是两个NRL撰写一章，另外一个NRL审核一章。结论HPV实验室电子手册是朝着全球统一HPV检测实验室方法迈出的一步，为研究和宫颈癌控制工作奠定了基础。

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引用次数: 0

Epidemiology and genotypic diversity of rhinovirus in school-age children with acute respiratory illnesses seeking medical care 求医急性呼吸道疾病学龄儿童鼻病毒流行病学及基因型多样性

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-08-01 Epub Date: 2025-05-28 DOI: 10.1016/j.jcv.2025.105806

Dithi Banerjee , Jennifer E. Schuster , Claire M. Midgley , Brian Lee , Mary Moffatt , Joana Y. Lively , Ariana P. Toepfer , Geoffrey A. Weinberg , Julie A. Boom , Leila C. Sahni , Vasanthi Avadhanula , Pedro A. Piedra , Mary Allen Staat , Daniel C. Payne , Natasha Halasa , John V. Williams , Robert W. Hickey , Marian G. Michaels , Janet A. Englund , Eileen J. Klein , Rangaraj Selvarangan

Background

Rhinovirus (RV) associated acute respiratory illness (ARI) data come mostly from infants and young children. We present data from 5 to 17-year-olds to characterize RV species A, B and C.

Methods

During December 1, 2016–Nov 30, 2017, seven U.S. New Vaccine Surveillance Network (NVSN) sites performed active pediatric ARI surveillance of inpatients (IP) and emergency department (ED) patients using molecular platforms to detect multiple respiratory pathogens. RV or RV/enterovirus (EV) positive specimens without co-detections were sequenced. Demographic and clinical data collected via parent interview and chart review were analyzed descriptively by RV species, month, and hospital setting, using chi-squared tests for comparisons.

Results

RV or RV/EV was detected in 581/2298 (25.3 %) ARI patients; 529 were single detections, 516 of these had sufficient sample for sequencing, and 420 (81.4 %) yielded sequence results: RV-A (183, 35.5 %), RV-B (16, 3.1 %), RV-C (210, 40.7 %), non-typeable RV (2, 0.4 %), and EV (9, 1.7 %). Among 52 RV-A, 8 RV-B and 44 RV-C unique types identified, A49 (32, 61.5 %), B6 (5, 62.5 %) and C15 (22, 50 %) were predominant. History of asthma was reported in 65.4 % RV-A, 50 % RV-B and 78.5 % RV-C patients (p = 0.005). Hospitalization occurred in 63.4 % RV-A, 37.5 % RV-B and 71.4 % RV-C patients (p = 0.012). RV-C detections peaked during winter and RV-A peaked during summer-fall.

Conclusions

RV exhibited genetic diversity in 5–17-year-old ARI patients, and circulation differed by RV species. Among children seeking care in ED or hospital settings, with confirmed RV or RV/EV single detections, hospitalization was more common with RV-A and -C than RV-B.

背景：鼻病毒（RV）相关的急性呼吸道疾病（ARI）数据主要来自婴幼儿。方法在2016年12月1日至2017年11月30日期间，美国新疫苗监测网络（NVSN）的7个站点使用分子平台检测多种呼吸道病原体，对住院患者（IP）和急诊科（ED）患者进行了儿科ARI监测。对未合并检测的RV或RV/ EV阳性标本进行测序。通过父母访谈和图表回顾收集的人口统计学和临床数据按RV品种、月份和医院设置进行描述性分析，使用卡方检验进行比较。结果2298例急性呼吸道感染患者中有581例（25.3%）检出RV或RV/EV；529例为单次检测，其中516例具有足够的测序样本，420例（81.4%）获得测序结果：RV- a（183例，35.5%）、RV- b（16例，3.1%）、RV- c（210例，40.7%）、不可分型RV（2例，0.4%）和EV（9例，1.7%）。在鉴定出的52株RV-A、8株RV-B和44株RV-C中，以A49（32, 61.5%）、B6（5, 62.5%）和C15（22, 50%）为主。65.4%的RV-A、50%的RV-B和78.5%的RV-C患者有哮喘病史（p = 0.005）。RV-A患者住院率为63.4%，RV-B患者为37.5%，RV-C患者为71.4% （p = 0.012）。RV-C检测在冬季达到高峰，RV-A检测在夏秋季达到高峰。结论5 ~ 17岁ARI患者srv具有遗传多样性，不同RV种类的循环存在差异。在急诊科或医院就诊的儿童中，确诊RV或RV/EV单一检测，RV- a和-C比RV- b更常见。

{"title":"Epidemiology and genotypic diversity of rhinovirus in school-age children with acute respiratory illnesses seeking medical care","authors":"Dithi Banerjee , Jennifer E. Schuster , Claire M. Midgley , Brian Lee , Mary Moffatt , Joana Y. Lively , Ariana P. Toepfer , Geoffrey A. Weinberg , Julie A. Boom , Leila C. Sahni , Vasanthi Avadhanula , Pedro A. Piedra , Mary Allen Staat , Daniel C. Payne , Natasha Halasa , John V. Williams , Robert W. Hickey , Marian G. Michaels , Janet A. Englund , Eileen J. Klein , Rangaraj Selvarangan","doi":"10.1016/j.jcv.2025.105806","DOIUrl":"10.1016/j.jcv.2025.105806","url":null,"abstract":"<div><h3>Background</h3><div>Rhinovirus (RV) associated acute respiratory illness (ARI) data come mostly from infants and young children. We present data from 5 to 17-year-olds to characterize RV species A, B and C.</div></div><div><h3>Methods</h3><div>During December 1, 2016–Nov 30, 2017, seven U.S. New Vaccine Surveillance Network (NVSN) sites performed active pediatric ARI surveillance of inpatients (IP) and emergency department (ED) patients using molecular platforms to detect multiple respiratory pathogens. RV or RV/enterovirus (EV) positive specimens without co-detections were sequenced. Demographic and clinical data collected via parent interview and chart review were analyzed descriptively by RV species, month, and hospital setting, using chi-squared tests for comparisons.</div></div><div><h3>Results</h3><div>RV or RV/EV was detected in 581/2298 (25.3 %) ARI patients; 529 were single detections, 516 of these had sufficient sample for sequencing, and 420 (81.4 %) yielded sequence results: RV-A (183, 35.5 %), RV-B (16, 3.1 %), RV-C (210, 40.7 %), non-typeable RV (2, 0.4 %), and EV (9, 1.7 %). Among 52 RV-A, 8 RV-B and 44 RV-C unique types identified, A49 (32, 61.5 %), B6 (5, 62.5 %) and C15 (22, 50 %) were predominant. History of asthma was reported in 65.4 % RV-A, 50 % RV-B and 78.5 % RV-C patients (<em>p</em> = 0.005). Hospitalization occurred in 63.4 % RV-A, 37.5 % RV-B and 71.4 % RV-C patients (<em>p</em> = 0.012). RV-C detections peaked during winter and RV-A peaked during summer-fall.</div></div><div><h3>Conclusions</h3><div>RV exhibited genetic diversity in 5–17-year-old ARI patients, and circulation differed by RV species. Among children seeking care in ED or hospital settings, with confirmed RV or RV/EV single detections, hospitalization was more common with RV-A and -C than RV-B.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"179 ","pages":"Article 105806"},"PeriodicalIF":4.0,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144221011","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}

引用次数: 0

Universal screening for congenital Cytomegalovirus infection using saliva from the neonate 利用新生儿唾液对先天性巨细胞病毒感染进行普遍筛查

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-06-01 Epub Date: 2025-04-20 DOI: 10.1016/j.jcv.2025.105800

Gabriele Halwachs-Baumann , Oliver Wagner , Yarub Salaheddin , Eveline Ziebermayr , Lukas Angleitner-Boubenizek , Georg Grüßenberger , Ulla Folger-Buchegger

Background

Congenital Cytomegalovirus infection (cCMV) is the most common intrauterine infection in developed countries. The diagnostic gold standard is detection of CMV in neonatal urine. Considering difficulties in urine sample collection, saliva is an alternative specimen providing supportive material for early diagnosis and timely intervention.

Objectives

Aim of this study was to evaluate a universal screening program using saliva as target specimen, and urine as confirmatory material.

Results

Between May 2019 and November 2024, 5168 newborns were screened for CMV shedding in saliva, 45 neonates were tested positive. From 44 neonates a urine sample was examined. Twelve out of forty-four neonates had concordant results (prevalence = 0,232 %) with five newborns showing CMV related symptoms. In saliva, the difference between the Ct-value of the cCMV negative and the cCMV positive group was significant (38.59 vs. 28.56, p-value < 0.0001). At a Ct-value cut-off of 38.8 sensitivity was 100 %, specificity 53.1 %, positive predictive value 44 %, and negative predictive value 100 %. ROC analysis yielded an AUC value of 0.930. The mean virus load in urine was 31,266,663 copies/ml.

Discussion

This study reports a prevalence of 0,232 % for cCMV infection in the catchment area of the hospital Steyr. PCR based CMV detection in saliva is a good screening tool, but one must be aware of false positive results, arising from possible contamination. In our cohort CMV positive vaginal or cervical secretion seems more likely being the source of contamination than CMV positive breast milk. The high false positive rate in saliva makes confirmatory testing in urine samples mandatory.

先天性巨细胞病毒感染（cCMV）是发达国家最常见的宫内感染。诊断的金标准是新生儿尿液中巨细胞病毒的检测。考虑到尿液样本采集的困难，唾液是一种可替代的样本，为早期诊断和及时干预提供支持材料。目的探讨一种以唾液为目标样本，尿液为确认材料的通用筛查方案。结果2019年5月至2024年11月，对5168名新生儿进行唾液巨细胞病毒脱落筛查，45名新生儿检测呈阳性。对44名新生儿进行了尿样检查。44名新生儿中有12名结果一致（患病率= 0,232%），其中5名新生儿出现巨细胞病毒相关症状。在唾液中，cCMV阴性组和cCMV阳性组的ct值差异有统计学意义(38.59比28.56，p值<；0.0001)。ct值为38.8时，敏感性为100%，特异性为53.1%，阳性预测值为44%，阴性预测值为100%。ROC分析的AUC值为0.930。尿中平均病毒载量为31,266,663拷贝/ml。本研究报告了斯太尔医院集水区cCMV感染率为0,232%。基于PCR的唾液巨细胞病毒检测是一个很好的筛选工具，但必须注意假阳性结果，可能引起污染。在我们的队列中，巨细胞病毒阳性的阴道或宫颈分泌物似乎比巨细胞病毒阳性的母乳更可能是污染源。唾液的高假阳性率使得尿样的确证测试成为强制性的。

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引用次数: 0

Use of polyethylene glycol precipitation and ultracentrifugation to enhance the sensitivity of hepatitis B virus DNA detection 采用聚乙二醇沉淀法和超离心法提高乙型肝炎病毒DNA检测的灵敏度

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-06-01 Epub Date: 2025-05-03 DOI: 10.1016/j.jcv.2025.105802

Michael X. Fu , Osmany Larralde , Richard Mayne , Kai Kean , Kaitlin Reid , Monique Andersson , Tanya Golubchik , Jane A. McKeating , Lisa Jarvis , William L. Irving , Peter Simmonds , Heli Harvala

Background

Sensitive molecular detection of hepatitis B virus (HBV) DNA is crucial for diagnosing and managing occult hepatitis. To improve the sensitivity of HBV DNA detection, we compared the effectiveness of polyethylene glycol (PEG) precipitation and ultracentrifugation to concentrate DNA prior to extraction.

Methods

Twenty-three HBV DNA-positive samples with low viral loads were compared between the extraction of standard (0.2 mL) and larger volumes (5 mL) of plasma, through PEG precipitation of 10 mL and 20 mL of plasma, and ultracentrifugation from 35 mL of plasma. The effectiveness of the methods for HBV DNA detection was assayed by quantitative PCR. For genetic characterisation, Sanger sequencing of amplicons and targeted Illumina sequencing were used. Costs, sample capacities, and turnaround times were compared.

Results

DNA was detected in a greater number of samples using PEG and ultracentrifugation (detecting up to all 23 samples) compared to more standard extraction methods (detecting at least 18 samples). Efficiencies of recovery of HBV DNA from samples were comparable in all concentration methods. HBV and other DNA viruses, such as human herpesviruses and anelloviruses, were detected in samples and at higher read counts with PEG concentration than without. The availability, cost, relative simplicity, and throughput of PEG precipitation conferred further advantages to ultracentrifugation.

Conclusions

PEG precipitation from large volumes of plasma is a practical and economical alternative to ultracentrifugation and could be a similarly effective concentration method for low viral load samples in blood donation and clinical virology laboratories.

背景：乙型肝炎病毒（HBV） DNA的敏感分子检测对于诊断和治疗隐匿性肝炎至关重要。为了提高HBV DNA检测的灵敏度，我们比较了聚乙二醇（PEG）沉淀和超离心在提取前浓缩DNA的有效性。方法对23例低病毒载量HBV dna阳性样本，分别采用标准血浆（0.2 mL）和大容量血浆（5 mL）、PEG沉淀血浆（10 mL和20 mL）、超离心血浆（35 mL）进行比较。采用定量PCR方法检测HBV DNA的有效性。为了进行遗传表征，扩增子采用Sanger测序和靶向Illumina测序。比较了成本、样品容量和周转时间。结果与更多标准提取方法（检测至少18个样品）相比，使用PEG和超离心（检测最多23个样品）在更多样品中检测到dna。从样品中回收HBV DNA的效率在所有浓度方法中都是相当的。在样品中检测到HBV和其他DNA病毒，如人类疱疹病毒和无绒病毒，并且在PEG浓度下检测到更高的读取计数。聚乙二醇沉淀的可用性、成本、相对简单性和吞吐量赋予了超离心进一步的优势。结论从大量血浆中提取speg沉淀是一种实用且经济的超离心方法，对于献血和临床病毒学实验室的低病毒载量样品是一种同样有效的浓缩方法。

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引用次数: 0

Isolation of Human Metapneumovirus from clinical specimen in human organoid-derived bronchial cell cultures is superior to isolation in monolayer cell line cultures 从人类器官支气管细胞培养的临床标本中分离人偏肺病毒优于从单层细胞系培养中分离

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-06-01 Epub Date: 2025-05-15 DOI: 10.1016/j.jcv.2025.105805

Pau Ribó-Molina, Stefan van Nieuwkoop, Mathis Funk , Babs E. Verstrepen, Jeroen J.A. van Kampen, Ron A.M. Fouchier, Bernadette G. van den Hoogen

Background

Human Metapneumovirus (HMPV) is a causative agent of respiratory tract infections (RTI) in children and adults. HMPV is a member of the Pneumoviridae family for which circulation of two serotypes, A and B, has been reported. HMPV isolation in standard monolayer cell lines is not always successful. Recently, it was shown that upon inoculation of human organoid-derived bronchial (ODB) cultures, HMPV primarily targeted the ciliated cells, similar as observed in experimentally infected animals. These observations lead to the hypothesis that isolation of virus from clinical specimen in this ODB model could be more successful than in standard monolayer cultures.

Methods

This study compared the efficiency of isolation of HMPV from 36 clinical samples in human ODB cultures with that in monolayers of Vero-118 cells.

Results

A total of 27 isolates (8 HMPV A and 19 HMPV B) were obtained in the ODB cultures, after one passage, whereas 21 isolates (9 HMPV A and 12 HMPV B) were obtained after one or two passages in Vero-118 cells.

Conclusions

Overall, the isolation efficiency of serotype A HMPV was comparable in both models, while isolation of serotype B viruses was profoundly more efficient in the ODB cultures than in Vero-118 cells, suggesting that primary cultures expressing ciliated cells should be considered as a superior isolation method for HMPV from clinical specimens.

背景：人偏肺病毒（HMPV）是儿童和成人呼吸道感染（RTI）的病原体。HMPV是肺炎病毒科的一员，已报告有两种血清型a和B的传播。在标准单层细胞系中分离HMPV并不总是成功的。最近，研究表明，接种人类器官衍生支气管（ODB）培养物后，HMPV主要针对纤毛细胞，这与在实验感染动物中观察到的情况相似。这些观察结果导致假设，在这种ODB模型中从临床标本中分离病毒可能比在标准单层培养中更成功。方法本研究比较了从36个临床标本中分离HMPV的效率和从Vero-118细胞单层中分离HMPV的效率。结果ODB培养1代共分离到HMPV A 8株、HMPV B 19株，而Vero-118细胞1、2代共分离到HMPV A 9株、HMPV B 12株。结论在两种模型中，血清A型HMPV的分离效率相当，而血清B型HMPV在ODB培养物中分离效率远高于在Vero-118细胞中分离效率，提示表达纤毛细胞的原代培养物可作为临床标本中分离HMPV的较优方法。

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引用次数: 0

Clinical and sociodemographic correlates of emergent or evolving HIV drug resistance in low viral load specimens in British Columbia, Canada 加拿大不列颠哥伦比亚省低病毒载量标本中出现或发展的HIV耐药性的临床和社会人口学相关性

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-06-01 Epub Date: 2025-05-28 DOI: 10.1016/j.jcv.2025.105807

Hanwei Sudderuddin , Charlotte Johanna Beelen , Jenny Li , Wendy Zhang , Melanie C.M. Murray , Viviane D. Lima , Julio S.G. Montaner , Chanson J. Brumme

Background/methods

Treatment guidelines recommend genotypic HIV drug resistance testing (DRT) at virologic failure, typically for plasma viral loads (pVL) > 1000 HIV RNA c/mL. In some settings, DRT can be performed on low viral load (LVL) samples (pVL of 50–250 c/mL); however, such testing is resource-intensive and its clinical benefit is unclear. Therefore, we investigated the frequency and factors associated with emergent resistance in LVL samples using a comprehensive, provincial database of HIV Protease-Reverse Transcriptase and Integrase sequences.

Results

A total of 43,979 Protease-RT DRTs were performed in British Columbia between 1999 and 2022, of which 2970 (6.8 %) were on LVL samples. Testing was successful for 1575 (53.0 %) LVL samples compared to 81.4 % and 94.4 % of samples with pVL 250–999 and pVL ≥ 1000 c/mL, respectively (p < 0.001). Compared to prior genotypes collected from samples with pVL > 250 c/mL, a total of 104 (7.3 %) cases of new or evolving drug resistance were identified from 1423 LVL DRTs. Of these, 49.5 %, 42.9 % and 22.9 % exhibited new Nucleoside Reverse Transcriptase, Non-Nucleoside Reverse Transcriptase and Protease resistance, respectively. Of 9309 Integrase DRTs performed between 2008 and 2022, only 4 (1.2 %) cases of new or evolving integrase resistance were observed. Multivariable analyses identified clinical/sociodemographic factors significantly associated with emergent resistance, including time elapsed between DRTs, historic cumulative resistance and NNRTI-based antiretroviral therapy.

Conclusions

Emergent or evolving resistance is identified infrequently in low viral load specimens. Given its resource-intensive nature, resistance testing of low viral load specimens may not be generally warranted.

背景/方法治疗指南建议在病毒学失败时进行基因型HIV耐药检测（DRT），通常用于检测血浆病毒载量（pVL） >；1000 HIV RNA c/mL。在某些情况下，DRT可以在低病毒载量（LVL）样品（pVL为50-250 c/mL）上进行；然而，这种检测是资源密集型的，其临床效益尚不清楚。因此，我们利用一个综合性的省级HIV蛋白酶-逆转录酶和整合酶序列数据库，调查了LVL样本中出现耐药性的频率和相关因素。结果1999年至2022年，不列颠哥伦比亚省共进行了43,979例蛋白酶rt DRTs，其中LVL样本为2970例（6.8%）。1575个（53.0%）LVL样品检测成功，而pVL 250-999和pVL≥1000 c/mL的样品分别为81.4%和94.4% (p <；0.001)。与先前从pVL和gt样本中收集的基因型进行比较；从1423例LVL DRTs中共鉴定出104例（7.3%）新的或正在发展的耐药病例。其中，49.5%、42.9%和22.9%分别表现出新的核苷逆转录酶、非核苷逆转录酶和蛋白酶抗性。在2008年至2022年期间进行的9309例整合酶drt中，仅观察到4例（1.2%）新的或进化的整合酶耐药病例。多变量分析确定了与出现耐药性显著相关的临床/社会人口因素，包括drt、历史累积耐药性和基于nnrti的抗逆转录病毒治疗之间的时间间隔。结论在低病毒载量标本中，出现耐药性或进化耐药性的情况并不多见。鉴于其资源密集型性质，通常不需要对低病毒载量标本进行耐药性检测。

{"title":"Clinical and sociodemographic correlates of emergent or evolving HIV drug resistance in low viral load specimens in British Columbia, Canada","authors":"Hanwei Sudderuddin , Charlotte Johanna Beelen , Jenny Li , Wendy Zhang , Melanie C.M. Murray , Viviane D. Lima , Julio S.G. Montaner , Chanson J. Brumme","doi":"10.1016/j.jcv.2025.105807","DOIUrl":"10.1016/j.jcv.2025.105807","url":null,"abstract":"<div><h3>Background/methods</h3><div>Treatment guidelines recommend genotypic HIV drug resistance testing (DRT) at virologic failure, typically for plasma viral loads (pVL) > 1000 HIV RNA c/mL. In some settings, DRT can be performed on low viral load (LVL) samples (pVL of 50–250 c/mL); however, such testing is resource-intensive and its clinical benefit is unclear. Therefore, we investigated the frequency and factors associated with emergent resistance in LVL samples using a comprehensive, provincial database of HIV Protease-Reverse Transcriptase and Integrase sequences.</div></div><div><h3>Results</h3><div>A total of 43,979 Protease-RT DRTs were performed in British Columbia between 1999 and 2022, of which 2970 (6.8 %) were on LVL samples. Testing was successful for 1575 (53.0 %) LVL samples compared to 81.4 % and 94.4 % of samples with pVL 250–999 and pVL ≥ 1000 c/mL, respectively (p < 0.001). Compared to prior genotypes collected from samples with pVL > 250 c/mL, a total of 104 (7.3 %) cases of new or evolving drug resistance were identified from 1423 LVL DRTs. Of these, 49.5 %, 42.9 % and 22.9 % exhibited new Nucleoside Reverse Transcriptase, Non-Nucleoside Reverse Transcriptase and Protease resistance, respectively. Of 9309 Integrase DRTs performed between 2008 and 2022, only 4 (1.2 %) cases of new or evolving integrase resistance were observed. Multivariable analyses identified clinical/sociodemographic factors significantly associated with emergent resistance, including time elapsed between DRTs, historic cumulative resistance and NNRTI-based antiretroviral therapy.</div></div><div><h3>Conclusions</h3><div>Emergent or evolving resistance is identified infrequently in low viral load specimens. Given its resource-intensive nature, resistance testing of low viral load specimens may not be generally warranted.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"178 ","pages":"Article 105807"},"PeriodicalIF":4.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144189834","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}

引用次数: 0

Comparison of mid-turbinate nasal and combined nasal-throat specimen types for detection of respiratory viruses in children 儿童中鼻甲鼻与合并鼻咽标本类型检测呼吸道病毒的比较

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology

Pub Date : 2025-06-01 Epub Date: 2025-04-29 DOI: 10.1016/j.jcv.2025.105801

Irem Onalan , Zheyi Teoh , Sarah L. Steele , Eileen J. Klein , Bonnie Strelitz , Kirsten Lacombe , Erin M. Sullivan , Arun K. Nalla , Danielle M. Zerr , Janet A. Englund

Background

The source of respiratory specimens may impact the detection of respiratory viruses. It may also have implications for research participant recruitment, caregiver acceptability due to concerns for patient comfort, and potential risk of aerosolization during epidemics/pandemics.

Objective

To determine the impact of collecting a throat swab (TS) in addition to a mid-turbinate nasal swab (MTS) by comparing agreement of viral detection and relative viral loads.

Study design

We reviewed molecular detection results from 2548 children enrolled in the New Vaccine Surveillance Network at Seattle Children’s Hospital from 11/2015–05/2019. Participants with a clinical MTS who agreed to collection of a combined TS and MTS (TS&MTS) for research were included. All specimens were tested using FilmArray^R Respiratory Panel (Biofire Diagnostics). Viral detection from MTS and TS&MTS were compared. Relative viral loads were compared between specimens with concordant (same viruses detected) and discordant (different or additional viruses detected) results.

Results

Results from 743 participants with clinical MTS and research TS&MTS specimens were compared; Viral detections were similar between the two groups, including 596 (80.2 %) paired results that were concordant. The most common discordant viruses were rhinovirus/enterovirus, respiratory syncytial virus, and adenovirus. Mean relative viral loads were lower in discordant specimens compared to concordant specimens, regardless of specimen source.

Conclusion

Comparison of clinical and research specimens revealed that respiratory viral detection was similar with or without an added TS. Lower relative viral loads of discordant specimens suggest that a combined TS&MTS may not improve viral detection for clinically significant pathogens.

呼吸道标本的来源可能影响呼吸道病毒的检测。它还可能对研究参与者的招募、护理人员对患者舒适度的接受程度以及流行病/大流行期间雾化的潜在风险产生影响。目的通过比较病毒检测结果与相对病毒载量的一致性，确定除中鼻甲鼻拭子外还采集咽拭子（TS）的影响。我们回顾了2015年11月至2019年5月在西雅图儿童医院新疫苗监测网络中登记的2548名儿童的分子检测结果。患有临床MTS的参与者同意收集TS和MTS （TS&；MTS）联合用于研究。所有标本均使用FilmArrayR呼吸面板（Biofire Diagnostics）进行检测。比较MTS和TS&；MTS的病毒检测结果。比较结果一致（检测到相同病毒）和不一致（检测到不同或额外的病毒）的标本之间的相对病毒载量。结果对743例临床MTS和研究MTS标本的结果进行了比较；两组之间的病毒检测结果相似，其中596（80.2%）对结果一致。最常见的不一致病毒是鼻病毒/肠病毒、呼吸道合胞病毒和腺病毒。不一致标本的平均相对病毒载量低于一致标本，无论标本来源如何。结论临床标本与研究标本的比较表明，添加或不添加TS的呼吸道病毒检出率相似，不一致标本的相对病毒载量较低，表明联合使用TS和MTS可能不能提高对临床重要病原体的病毒检出率。

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引用次数: 0