A universal RT-qPCR assay for "One Health" detection of influenza A viruses.
A universal RT-qPCR assay for "One Health" detection of influenza A viruses.
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DOI：
10.1371/journal.pone.0244669
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Brown IH
中科院分区：
文献类型：
--
作者：
Nagy A;Černíková L;Kunteová K;Dirbáková Z;Thomas SS;Slomka MJ;Dán Á;Varga T;Máté M;Jiřincová H;Brown IH
人类和动物健康的相互依赖是“同一个健康”倡议作为传染病控制和管理综合战略的核心。 “同一个健康”的一个关键要素包括准备和应对人与动物界面上的甲型流感病毒 (IAV) 威胁。 IAV 的特点是广泛的遗传变异性，它们在不同宿主之间循环，并可以建立宿主特异性谱系。四种主要宿主是：禽类、猪、人和马，偶尔传播给其他哺乳动物物种。宿主多样性反映在 IAV 检测的 RT-qPCR 检测范围内。卫生主管部门建议采用不同的检测方法对鸟类、猪或人类进行通用 IAV 检测。为了统一不同宿主的 IAV 监测并应用 One Health 方法，我们开发了一种单一 RT-qPCR 检测方法，用于对所有亚型、物种起源和全球分布的所有 IAV 进行通用检测。该检测设计以 IAV 基质蛋白 (MP) 片段的高度保守区域为中心，该区域是通过对 99,353 个序列的综合分析而确定的。反应参数得到有效优化，每个反应约 10 个 IAV 模板的效率为 93-97%，LOD 为 95%。该测定显示出高重复性、再现性和稳健性。广泛的计算机评估显示了高包容性，即引物和探针结合区域的完美序列匹配，猪的匹配率为 94.6%，禽类的匹配率为 98.2%，人 H3N2、大流行性 H1N1 以及其他 IAV 毒株的匹配率为 100%，分析数据集的总体预测检出率为 99%。六个兽医或人类诊断实验室合作对总共 1970 个样本进行了确认和广泛验证，其中 1455 个是临床样本，包括多种 IAV 毒株。
The mutual dependence of human and animal health is central to the One Health initiative as an integrated strategy for infectious disease control and management. A crucial element of the One Health includes preparation and response to influenza A virus (IAV) threats at the human-animal interface. The IAVs are characterized by extensive genetic variability, they circulate among different hosts and can establish host-specific lineages. The four main hosts are: avian, swine, human and equine, with occasional transmission to other mammalian species. The host diversity is mirrored in the range of the RT-qPCR assays for IAV detection. Different assays are recommended by the responsible health authorities for generic IAV detection in birds, swine or humans. In order to unify IAV monitoring in different hosts and apply the One Health approach, we developed a single RT-qPCR assay for universal detection of all IAVs of all subtypes, species origin and global distribution. The assay design was centred on a highly conserved region of the IAV matrix protein (MP)-segment identified by a comprehensive analysis of 99,353 sequences. The reaction parameters were effectively optimised with efficiency of 93–97% and LOD95% of approximately ten IAV templates per reaction. The assay showed high repeatability, reproducibility and robustness. The extensive in silico evaluation demonstrated high inclusivity, i.e. perfect sequence match in the primers and probe binding regions, established as 94.6% for swine, 98.2% for avian and 100% for human H3N2, pandemic H1N1, as well as other IAV strains, resulting in an overall predicted detection rate of 99% on the analysed dataset. The theoretical predictions were confirmed and extensively validated by collaboration between six veterinary or human diagnostic laboratories on a total of 1970 specimens, of which 1455 were clinical and included a diverse panel of IAV strains.
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DOI：
10.1093/emph/eoy013
发表时间：
2018
期刊：
Evolution, medicine, and public health
影响因子：
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作者：
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影响因子：
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DOI：
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发表时间：
2018-06-28
期刊：
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影响因子：
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作者：
Krammer F;Smith GJD;Fouchier RAM;Peiris M;Kedzierska K;Doherty PC;Palese P;Shaw ML;Treanor J;Webster RG;García-Sastre A
通讯作者：
García-Sastre A
影响因子：
4.8
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Heine HG;Foord AJ;Wang J;Valdeter S;Walker S;Morrissy C;Wong FY;Meehan B
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DOI：
10.1071/nb11005
发表时间：
2011-07-01
期刊：
New South Wales public health bulletin
影响因子：
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作者：
Dwyer, Dominic E;Kirkland, Peter D
通讯作者：
Kirkland, Peter D