Development of a TaqMan MGB RT-PCR for the rapid detection of H3 subtype avian influenza virus circulating in China

文献类型： 外文期刊

第一作者： Teng, Qiaoyang

作者： Teng, Qiaoyang;Shen, Weixia;Yan, Dawei;Yan, Liping;Li, Xuesong;Li, Guoxin;Yang, Jianmei;Li, Zejun;Teng, Qiaoyang;Yan, Liping;Li, Xuesong;Li, Guoxin;Yang, Jianmei;Li, Zejun

作者机构：

关键词： Minor groove binder;H3 subtype;Sensitivity;Detection

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Previous studies demonstrated that the H3 avian influenza virus (AIV) in China is isolated most frequently from wild birds and live poultry markets. However, there is no subtype-specific real-time polymerase chain reaction (RT-PCR) available for the rapid and highly sensitive identification of H3 AIV. In this study, a TaqMan minor groove binder (MGB) probe and a pair of primers were designed based on a conserved region in the hemagglutinin gene of H3 AIV. These were used to generate an H3-MGB RT-PCR assay that recognizes only H3 AIV. The detection limit of the H3-MGB RT-PCR was 10 copies of DNA per reaction when 10-fold serial dilutions of T-H3HA plasmid were used as the template. This was 1000-times more sensitive than conventional RT-PCR. In experimental samples obtained from oropharyngeal swabs or cloacal swabs, the virus was detected in all ducks using H3-MGB RT-PCR, whereas only one duck tested positive for the virus in oropharyngeal swabs tested using conventional RT-PCR. The H3-MGB RT-PCR assay developed in this study is a sensitive and rapid tool for screening H3 AIV in China. (C) 2015 Elsevier B.V. All rights reserved.

分类号： R37

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