Genetic and Molecular Characterization of H9N2 Avian Influenza Viruses Isolated from Live Poultry Markets in Hubei Provi
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RESEARCH ARTICLE
Genetic and Molecular Characterization of H9N2 Avian Influenza Viruses Isolated from Live Poultry Markets in Hubei Province, Central China, 2013–2017 Zhibin Hu1,2 • Fuhu Peng2 • Zhenghui Xiong2 • Wanpo Zhang1 • Tingting Li2 • Yuejun Shi1 • Jun Xie2 Xin Jin2 • Jingjing Huang2 • Hongde Xiao2 • Dingren Bi1 • Nianhua Song2 • Zili Li1
•
Received: 17 January 2020 / Accepted: 24 April 2020 Ó Wuhan Institute of Virology, CAS 2020
Abstract H9N2 subtype avian influenza virus (AIV) is an influenza A virus that is widely spread throughout Asia, where it jeopardizes the poultry industry and provides genetic material for emerging human pathogens. To better understand the epidemicity and genetics of H9 subtype AIVs, we conducted active surveillance in live poultry markets (LPMs) in Hubei Province from 2013 to 2017. A total of 4798 samples were collected from apparent healthy poultry and environment. Realtime RT-PCR revealed that the positivity rate of influenza A was 26.6% (1275/4798), of which the H9 subtype accounted for 50.3% (641/1275) of the positive samples. Of the 132 H9N2 viral strains isolated, 48 representative strains were subjected to evolutionary analysis and genotyping. Phylogenetic analysis revealed that all H9N2 viral genes had 91.1%– 100% nucleotide homology, clustered with genotype 57, and had high homology with human H9N2 viruses isolated from 2013 to 2017 in China. Using a nucleotide divergence cutoff of 95%, we identified ten distinct H9N2 genotypes that continued to change over time. Molecular analysis demonstrated that six H9N2 isolates had additional potential glycosylation sites at position 218 in the hemagglutinin protein, and all isolates had I155T and Q226L mutations. Moreover, 44 strains had A558V mutations in the PB2 protein and four had E627V mutations, along with H9N2 human infection strains A/Beijing/1/2016 and A/Beijing/1/2017. These results emphasize that the H9N2 influenza virus in Hubei continues to mutate and undergo mammalian adaptation changes, indicating the necessity of strengthening the surveillance of the AIV H9N2 subtype in LPMs. Keywords Avian influenza H9N2 Genetic variation Phylogenetic analysis Live poultry markets (LPMs)
Introduction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12250-020-00260-z) contains supplementary material, which is available to authorized users. & Zili Li [email protected] & Nianhua Song [email protected] 1
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
2
Hubei Center for Animal Disease Control and Prevention, Wuhan 430070, China
The influenza A virus has eight distinct gene segments that can be divided into 18 hemagglutinin (HA) and 11 neuraminidase (NA) subtypes, of which H1–H16 are found in birds and H17 and H18 are found in bats (Webster et al. 1992; Tong et al. 2013). H9N2, H5Nx, and H7N9 subtype influenza A viruses cause the most harm to the pou
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