迈向诊断准备:利用快速抗原和即时分子检测在人造鼻拭子标本中检测高致病性甲型H5N1禽流感。

IF 5.4 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-09-10 Epub Date: 2025-08-19 DOI:10.1128/jcm.00548-25
Leda Bassit, Gregory L Damhorst, Heather B Bowers, Courtney Sabino, Julie Sullivan, Emily B Kennedy, Jacob Khouri, Pamela Miller, Eric Lai, Raymond F Schinazi, Wilbur A Lam, Nira R Pollock, Anuradha Rao
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引用次数: 0

摘要

高致病性禽流感(HPAI) A(H5N1)分支2.3.4.4b于2021年在美国首次在鸟类中发现。奶牛中的持续疫情始于2024年初,涉及基因型B3.13。在美国,已发现至少70例与受感染的牛、家禽和野鸟接触的人感染病例。尚未有人际传播的记录。然而,在诊断准备方面,我们评估了目前可用的流感检测方法检测2024年美国H5N1基因型(B3.13和D1.1)的能力,这些基因型主要存在于野生鸟类和家禽中。使用活病毒或灭活病毒制备人造鼻拭子样本,用于检测12种快速抗原检测(侧流试验[LFA]),包括10种市售甲型流感和2种h5特异性LFA。还检测了5种即时护理(POC)分子测定法。使用预先确定的稀释系列的包容性测试方案来评估每个分析,从而实现性能的头对头比较。所有LFAs和POC分子检测均能检测到牛2024型H5N1基因型B3.13。POC分子测试(热灭活B3.13)的灵敏度范围为1.55至7.75 TCID50/拭子。对于11/12个LFAs, 10个商业甲型流感和一个RUO H5检测,敏感性(活B3.13)范围为78至1550 TCID50/拭子。第12次LFA检出TCID50/mL输运介质77500例。4个lfa的检测证实了基因型D1.1的包容性。现有的流感测试可以在具有广泛分析灵敏度的人造样本中检测到2024年美国H5N1病毒株。在人际传播的情况下,需要确定临床表现和最佳样本类型。迄今为止,已报告了至少70例人感染高致病性甲型禽流感(HPAI) H5N1病例。尚未报告人与人之间的传播,但高致病性H5N1在动物和鸟类种群中持续传播。作为大流行防范工作的一部分,至关重要的是要了解能够检测季节性甲型流感的现有检测方法是否也能检测高致病性H5N1。我们的论文描述了第一项研究的结果,该研究评估和比较了10种市售甲型流感横向流动测定法(LFAs)、两种H5流感特异性LFAs和三家制造商的五种POC分子检测法检测美国牛中流行的H5N1病毒(基因型B3.13)的能力。我们发现,所有检测方法都能在人造鼻拭子标本中检测到高致病性H5N1病毒,分析灵敏度不同。我们还测试了LFA的一个子集(三个510k清除的COVID/流感多重检测和一个RUO h5特异性LFA),以确认它们能够检测从华盛顿州H5N1人类病例中分离出来的D1.1基因型(野生鸟类和家禽的主要基因型)毒株。鉴于高致病性H5N1病例的持续传播,至关重要的是要知道现有的检测方法能够分析地发现H5N1。如果观察到人际传播,应评估临床表现和最佳样本类型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Toward diagnostic preparedness: detection of highly pathogenic avian influenza A(H5N1) in contrived nasal swab specimens using rapid antigen and point-of-care molecular tests.

Toward diagnostic preparedness: detection of highly pathogenic avian influenza A(H5N1) in contrived nasal swab specimens using rapid antigen and point-of-care molecular tests.

Highly pathogenic avian influenza (HPAI) A(H5N1) clade 2.3.4.4b was first detected in birds in 2021 in the United States (U.S.). An ongoing outbreak in dairy cattle began in early 2024 involving genotype B3.13. At least 70 U.S. cases have been identified in humans with exposure to infected cattle, poultry, and wild birds. No human-to-human transmission has yet been documented. However, for diagnostic preparedness, we evaluated the ability of currently available influenza tests to detect 2024 U.S. H5N1 genotypes, B3.13 and D1.1, predominant in wild birds and poultry. Contrived nasal swab samples were prepared using live or inactivated virus and used to test 12 rapid antigen tests (lateral flow assays, [LFA]), including 10 commercially available influenza A and two H5-specific LFAs. Five point-of-care (POC) molecular assays were also tested. An inclusivity testing protocol utilizing a predetermined dilution series was used to evaluate each assay, enabling a head-to-head comparison of performances. All LFAs and POC molecular tests were able to detect bovine 2024 H5N1 genotype B3.13. Sensitivity for the POC molecular tests (heat-inactivated B3.13) ranged from 1.55 to 7.75 TCID50/swab. For 11/12 LFAs, 10 commercial influenza A, and an RUO H5 assay, the sensitivity (live B3.13) ranged from 78 to 1550 TCID50/swab. The 12th LFA detected 77,500 TCID50/mL transport media. Testing of four LFAs confirmed inclusivity for genotype D1.1. Available influenza tests can detect 2024 U.S. H5N1 strains in contrived samples with a wide range of analytical sensitivity. In the event of human-to-human transmission, clinical performance and optimal sample types would need to be established.IMPORTANCETo date, at least 70 human cases of highly pathogenic avian influenza A (HPAI) H5N1 have been reported. Human-to-human transmission has not yet been reported, but there is ongoing HPAI H5N1 spread within animal and bird populations. As part of pandemic preparedness, it is critical to know whether available tests capable of detecting seasonal influenza A can also detect HPAI H5N1. Our paper describes results from the first study to assess and compare the ability of 10 commercially available influenza A lateral flow assays (LFAs), two LFAs specific for H5 influenza, and five POC molecular tests from three manufacturers to detect the H5N1 virus circulating in U.S. cattle (genotype B3.13). We found that all tests detect HPAI H5N1 in contrived nasal swab specimens, with varying degrees of analytical sensitivity. We also tested a subset of LFAs (three 510k-cleared COVID/flu multiplex tests and an RUO H5-specific LFA) to confirm that they were able to detect a D1.1 genotype (the predominant genotype in wild birds and poultry) strain isolated from a human case of H5N1 in Washington State. Given the persistent circulation of HPAI H5N1 cases, it is critical to know that available tests can analytically detect H5N1. Clinical performance and optimal sample types should be evaluated if human-to-human transmission is observed.

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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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