Enhancing diagnostic preparedness for H5N1: a validation study of H5 single-plex assay and detection across multiple platforms.

IF 5.4 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-08-13 Epub Date: 2025-07-18 DOI:10.1128/jcm.00681-25
Yuan Chao Xue, Jennifer Bertsch, Kaylin Monacy, Carter Haynes, Natalie Williams-Bouyer, Barbara M Judy, Patrick C Newman, Thomas G Ksiazek, Lyudmyla V Marushchak, Gregory C Gray, Ping Ren
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引用次数: 0

Abstract

The increasing transmission of highly pathogenic avian influenza (HPAI) H5N1 from animals to humans underscores the urgent need for enhanced diagnostic capabilities in clinical microbiology laboratories. Although in silico analysis suggests that commercial multiplex respiratory panels can detect H5N1, these assays lack subtyping H5N1 capabilities, and their real-world performance remains largely unverified. In this study, we evaluated the limit of detection (LoD) for H5N1 using three commercial molecular diagnostic assays routinely employed at our institution: bioMérieux BioFire Respiratory 2.1 Panel, Cepheid Xpert Xpress CoV-2/Flu/RSV Plus, and Hologic Panther Fusion SARS-CoV-2/Flu A/B/RSV assays. All three reliably detected H5N1 were at low viral concentrations. To enable H5N1 detection, we also developed a real-time RT-PCR H5 single-plex assay on the Hologic Panther Fusion Open Access platform for reflex testing of influenza A-positive specimens. In conclusion, although current commercial assays lack influenza A H5 subtype differentiation, our validation data provide critical performance information. When integrated with a targeted H5 assay, these tools can enhance clinical decision-making and public health surveillance by reducing the risk of missed H5N1 infection cases.IMPORTANCEThis study addresses a growing public health concern: the spread of bird flu (H5N1) from animals to humans. Most hospital laboratories use commercial tests to detect respiratory viruses like the flu, but these tests cannot tell if someone has the specific and more dangerous H5N1 strain. To help solve this, we tested three commonly used diagnostic tools and found that they can detect H5N1 even at low levels. However, since they cannot identify the specific H5 subtype, we also developed and validated a follow-up test that runs on one of the existing laboratory high-throughput equipment. This test can confirm whether a patient infected with the flu has the H5N1 strain. By combining these tools, hospital laboratories can improve early detection of H5N1, support better patient care, and help public health officials respond more effectively to outbreaks.

加强对H5N1的诊断准备:一项跨多个平台的H5单一检测和检测的验证研究。
H5N1型高致病性禽流感(HPAI)从动物向人类的传播日益增加,这突出表明迫切需要加强临床微生物学实验室的诊断能力。尽管计算机分析表明,商业多重呼吸检测板可以检测H5N1,但这些检测方法缺乏H5N1亚型分型能力,而且它们在现实世界中的表现在很大程度上仍未经证实。在本研究中,我们使用本机构常规使用的三种商业分子诊断检测方法评估了H5N1的检出限(LoD): biomrieux BioFire Respiratory 2.1 Panel、Cepheid Xpert Xpress CoV-2/Flu/RSV Plus和Hologic Panther Fusion SARS-CoV-2/Flu A/B/RSV检测。所有三个可靠检测到的H5N1病毒浓度都很低。为了检测H5N1,我们还在Hologic Panther Fusion开放获取平台上开发了一种实时RT-PCR H5单组分检测方法,用于甲型流感阳性标本的反射检测。总之,尽管目前的商业检测缺乏甲型流感H5亚型分化,但我们的验证数据提供了关键的性能信息。当与有针对性的H5检测相结合时,这些工具可以通过减少漏诊H5N1感染病例的风险来加强临床决策和公共卫生监测。重要意义本研究解决了一个日益严重的公共卫生问题:禽流感(H5N1)从动物到人类的传播。大多数医院实验室使用商业测试来检测流感等呼吸道病毒,但这些测试无法判断某人是否感染了特定的、更危险的H5N1病毒株。为了帮助解决这个问题,我们测试了三种常用的诊断工具,发现它们即使在低水平也能检测到H5N1。然而,由于他们无法识别特定的H5亚型,我们还开发并验证了在现有实验室高通量设备上运行的后续测试。该测试可确认感染流感的患者是否携带H5N1毒株。通过结合这些工具,医院实验室可以改善H5N1的早期发现,支持更好的病人护理,并帮助公共卫生官员更有效地应对疫情。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
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