Rapid and sensitive detection of high pathogenicity Eurasian clade 2.3.4.4b avian influenza viruses in wild birds and poultry
Rapid and sensitive detection of high pathogenicity Eurasian clade 2.3.4.4b avian influenza viruses in wild birds and poultry
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DOI：
10.1016/j.jviromet.2022.114454
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发表时间：
2022-01-12
影响因子：
3.1
通讯作者：
Reid, Scott M.
中科院分区：
文献类型：
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作者：
James, Joe;Seekings, Amanda H.;Reid, Scott M.
禽流感病毒（Avian influenza virus，AIV）根据对鸡的静脉注射致病性和/或血凝素（Heamagglutinin，HA）裂解位点（cleavage site，CS）是否存在多个碱性残基，分为高致病性和低致病性AIV（HPAIV/LPAIV）。自2014年以来，欧洲经历了H5 Nx HPAIV的入侵浪潮。在2020年11月至2021年3月期间，这些包括HPAIV H5 N8，以及零星的H5 N1和H5 N5（所有分支2.3.4.4b），在英国通过被动监测计划提交的300多只“发现死亡”野鸟中检测到。目前，H5 Nx HPAIV检测依赖于使用实时逆转录PCR（rRT-PCR）测定来鉴定AIV RNA和H5亚型。随后通过HA CS的桑格测序确定致病型。在这里，我们报告的验证和应用的快速，更具成本效益的HP H5检测rRT-PCR检测。HP H5 rRT-PCR检测试剂盒特异性、灵敏度和可重复性地检测到来自当代进化枝2.3.4.4b H5 HPAIV的RNA，其灵敏度与诊断H5特异性rRT-PCR相当;未检测到LPAIV H5 RNA和非AIV RNA。对于来自“发现死亡”的野生鸟类的材料，以及家禽的法定疾病诊断，与常规CS测序相比，HP H5 rRT-PCR结果提供了100%的区分度，显著缩短了确定致病型的时间和成本，增强了诊断工作流程。
Avian influenza virus (AIV) is classified as high or low pathogenicity AIV (HPAIV/LPAIV) based on intravenous pathogenicity in chickens and/or the presence or absence of multiple basic residues at the heamagglutinin (HA) cleavage site (CS). Since 2014, Europe has experienced waves of incursions of H5Nx HPAIV. Between November 2020 and March 2021, these included HPAIV H5N8, with sporadic of H5N1 and H5N5 (all clade 2.3.4.4b), detected in more than 300 "found dead" wild birds submitted through a passive surveillance programme in the United Kingdom. Currently, H5Nx HPAIV detection relies on identification of AIV RNA and H5 subtyping using real-time reverse transcription PCR (rRT-PCR) assays. The pathotype is subsequently determined by Sanger sequencing of the HA CS. Here, we report the validation and application of a rapid, more cost-effective HP H5detection rRT-PCR assay. The HP H5 rRT-PCR assay specifically, sensitively and reproducibly detected RNA from contemporary clade 2.3.4.4b H5 HPAIVs with comparable sensitivity to the diagnostic H5-specific rRT-PCR; LPAIV H5 RNA and non-AIV RNA were not detected. On material from "found-dead" wild birds, and for statutory disease diagnosis on poultry, the HP H5 rRT-PCR results provided 100% discrimination when compared to conventional CS sequencing, significantly reducing time-to-pathotype determination and cost, enhancing the diagnostic workflow.