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首页> 外文期刊>Journal of Clinical Microbiology >Detection of Hemagglutinin Variants of the Pandemic Influenza A (H1N1) 2009 Virus by Pyrosequencing
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Detection of Hemagglutinin Variants of the Pandemic Influenza A (H1N1) 2009 Virus by Pyrosequencing

机译：焦磷酸测序法检测大流行性甲型流感（H1N1）2009病毒的血凝素变体

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摘要

For influenza viruses, pyrosequencing has been successfully applied to the high-throughput detection of resistance markers in genes encoding the drug-targeted M2 protein and neuraminidase. In this study, we expanded the utility of this assay to the detection of multiple receptor binding variants of the hemagglutinin protein of influenza viruses directly in clinical specimens. Specifically, a customized pyrosequencing protocol that permits detection of virus variants with the D, G, N, or E amino acid at position 222 in the hemagglutinin of the 2009 pandemic influenza A (H1N1) virus was developed. This customized pyrosequencing protocol was applied to the analysis of 241 clinical specimens. The use of the optimized nucleotide dispensation order allowed detection of mixtures of variants in 10 samples (4.1%) which the standard cyclic nucleotide dispensation protocol failed to detect. The optimized pyrosequencing protocol is expected to provide a more accurate tool in the analysis of virus variant composition.
机译：对于流行性感冒病毒，焦磷酸测序已成功应用于编码药物靶向的M2蛋白和神经氨酸酶的基因中抗性标记的高通量检测。在这项研究中，我们将该方法的实用性扩展到直接在临床标本中检测流感病毒血凝素蛋白的多个受体结合变异体。具体而言，开发了一种定制的焦磷酸测序方案，该方案允许检测2009年大流行性流感A（H1N1）病毒血凝素中222位D，G，N或E氨基酸的病毒变体。该定制的焦磷酸测序方案被用于分析241个临床标本。使用优化的核苷酸分配顺序可以检测10个样品中的变体混合物（4.1％），而标准环状核苷酸分配方案则无法检测到。经过优化的焦磷酸测序方案有望为病毒变体组成的分析提供更准确的工具。

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