甲型H1N1流感pdm09 6B.1A.5a的出现。a. a. a. a. a. a. a. b。在商业分子分析中，1个亚枝导致分型失败

IF 4 3区医学 Q2 VIROLOGY

Journal of Clinical Virology Pub Date : 2025-04-09 DOI:10.1016/j.jcv.2025.105797

Bosung Park, Eun Jeong Won, Heungsup Sung, Mi-Na Kim

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引用次数: 0

摘要

背景在2023-2024年和2024-2025年初的流感季节，我们实验室的一些甲型流感阳性标本未能通过Allplex Respiratory Panel 1 (Allplex RP1) (Seegene Inc.)进行H1、H1pdm09和H3亚型检测。材料与方法在 2023 年 8 月至 2024 年 12 月期间，23 份鼻咽标本的甲型流感检测结果呈阳性，但无法对 H1、H1pdm09 和 H3 进行亚型鉴定。制造商进行的确证检测包括 M 和 HA 基因的目标特异性 PCR，然后进行测序以确定亚型。其中，21 个属于 6B.1A.5a.2a.1 亚支系，1 个属于 6B.1A.5a.2a。序列分析表明，Allplex RP1 的 H1pdm09 引物/探针结合区存在错配，这解释了亚型划分失败的原因。结论6B.1A.5a.2a.1和6B.1A.5a.2a亚支系存在突变，导致一些用商业检测方法检测的样本亚型鉴定失败。虽然无法分型的甲型流感结果往往会引起人们对新出现菌株的担忧，但测序结果证实，用 Allplex RP1 检测的所有无法分型的样本都属于公认亚支系中的 H1N1pdm09。因此，尽管基因组监测仍然非常重要，但该检测方法的亚型鉴定失败并不一定表明存在新型病毒或人畜共患病毒。

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Emergence of influenza A(H1N1)pdm09 6B.1A.5a.2a and 6B.1A.5a.2a.1 subclades leading to subtyping failure in a commercial molecular assay

Background

During the 2023–2024 and early 2024–2025 influenza seasons, several influenza A-positive specimens in our laboratory failed subtyping for H1, H1pdm09, and H3 using the Allplex Respiratory Panel 1 (Allplex RP1) (Seegene Inc.). This study aimed to identify the cause of these subtyping failures.

Materials and methods

Between August 2023 and December 2024, 23 nasopharyngeal specimens tested positive for influenza A but were unsubtypeable for H1, H1pdm09, and H3. Confirmatory testing by the manufacturer included target-specific PCR for the M and HA genes, followed by sequencing to determine subclades.

Results

Among the 23 unsubtypeable specimens, 22 yielded PCR products for sequencing. Of these, 21 belonged to subclade 6B.1A.5a.2a.1 and one to 6B.1A.5a.2a. Sequence analysis revealed mismatches in the H1pdm09 primer/probe-binding regions of Allplex RP1, explaining the subtyping failures. Despite testing negative for H1pdm09 in Allplex RP1, sequencing confirmed their classification as H1N1pdm09 subclades with HA gene mutations.

Conclusions

Subclades 6B.1A.5a.2a.1 and 6B.1A.5a.2a harbour mutations that contributed to subtyping failures in some specimens tested with a commercial assay. While unsubtypeable influenza A results often raise concerns about emerging strains, sequencing confirmed that all unsubtypeable specimens tested with Allplex RP1 belonged to H1N1pdm09 within recognised subclades. Thus, such subtyping failures in this assay do not necessarily indicate a novel or zoonotic virus, though genomic surveillance remains essential.

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来源期刊

Journal of Clinical Virology 医学-病毒学

CiteScore

22.70

自引率

1.10%

发文量

149

审稿时长

24 days

期刊介绍： The Journal of Clinical Virology, an esteemed international publication, serves as the official journal for both the Pan American Society for Clinical Virology and The European Society for Clinical Virology. Dedicated to advancing the understanding of human virology in clinical settings, the Journal of Clinical Virology focuses on disseminating research papers and reviews pertaining to the clinical aspects of virology. Its scope encompasses articles discussing diagnostic methodologies and virus-induced clinical conditions, with an emphasis on practicality and relevance to clinical practice. The journal publishes on topics that include: • new diagnostic technologies • nucleic acid amplification and serologic testing • targeted and metagenomic next-generation sequencing • emerging pandemic viral threats • respiratory viruses • transplant viruses • chronic viral infections • cancer-associated viruses • gastrointestinal viruses • central nervous system viruses • one health (excludes animal health)