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Development of a real-time RT-PCR method for the detection of newly emerged highly pathogenic H7N9 influenza viruses

文献类型： 外文期刊

作者： Wang Xiu-rong 1 ; Gu Lin-lin 1 ; Shi Jian-zhong 1 ; Xu Hai-feng 1 ; Zhang Ying 1 ; Zeng Xian-ying 1 ; Deng Guo-hua 1 ; Li 1 ;

作者机构： 1.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Harbin 150069, Heilongjiang, Peoples R China

2.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol,

关键词： H7N9;highly pathogenic influenza virus;real-time RT-PCR

期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：2.848； 五年影响因子：2.979 ）

ISSN： 2095-3119

年卷期： 2017 年 16 卷 9 期

页码：

收录情况： SCI

摘要： In 2013, a human influenza outbreak caused by a novel H7N9 virus occurred in China. Recently, the H7N9 virus acquired multiple basic amino acids at its hemagglutinin (HA) cleavage site, leading to the emergence of a highly pathogenic virus. The development of an effective diagnostic method is imperative for the prevention and control of highly pathogenic H7N9 influenza. Here, we designed and synthesized three pairs of primers based on the nucleotide sequence at the HA cleavage site of the newly emerged highly pathogenic H7N9 influenza virus. One of the primer pairs and the corresponding probe displayed a high level of amplification efficiency on which a real-time RT-PCR method was established. Amplification using this method resulted in a fluorescent signal for only the highly pathogenic H7N9 virus, and not for any of the H1-H15 subtype reference strains, thus demonstrating high specificity. The method detected as low as 39.1 copies of HA-positive plasmid and exhibited similar sensitivity to the virus isolation method using embryonated chicken eggs. Importantly, the real-time RT-PCR method exhibited 100% consistency with the virus isolation method in the diagnosis of field samples. Collectively, our data demonstrate that this real-time RT-PCR assay is a rapid, sensitive and specific method, and the application will greatly aid the surveillance, prevention, and control of highly pathogenic H7N9 influenza viruses.

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