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Genetic and Molecular Characterization of H9N2 Avian Influenza Viruses Isolated from Live Poultry Markets in Hubei Province, Central China, 2013–2017

  • RESEARCH ARTICLE
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Virologica Sinica

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. Real-time 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.

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Acknowledgements

We would like to thank Dr. Yushan Chen from Wuhan University of Science and Technology for her help with data analysis. This study was supported by the National Key Research and Development Program of China (Grant No. 2016YFD0500800 to ZL).

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Authors and Affiliations

  1. College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China

    Zhibin Hu, Wanpo Zhang, Yuejun Shi, Dingren Bi & Zili Li

  2. Hubei Center for Animal Disease Control and Prevention, Wuhan, 430070, China

    Zhibin Hu, Fuhu Peng, Zhenghui Xiong, Tingting Li, Jun Xie, Xin Jin, Jingjing Huang, Hongde Xiao & Nianhua Song

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  1. Zhibin Hu
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  2. Fuhu Peng
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  3. Zhenghui Xiong
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  4. Wanpo Zhang
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  5. Tingting Li
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  6. Yuejun Shi
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  7. Jun Xie
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  8. Xin Jin
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  9. Jingjing Huang
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  10. Hongde Xiao
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  11. Dingren Bi
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  12. Nianhua Song
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  13. Zili Li
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Contributions

DRB and NS designed and coordinated the study. ZH, XJ, JX, JH, and HX collected samples. ZH, FP, and ZX carried out most of the experiments. ZH, TL, YS, and ZL analyzed the data. ZH, WZ, and ZL wrote and finalized the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Nianhua Song or Zili Li.

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The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

The study was approved by the Hubei Center for Animal Disease Control and Prevention. All institutional and national guidelines for the care and use of animals were followed.

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Hu, Z., Peng, F., Xiong, Z. et al. Genetic and Molecular Characterization of H9N2 Avian Influenza Viruses Isolated from Live Poultry Markets in Hubei Province, Central China, 2013–2017. Virol. Sin. 36, 291–299 (2021). https://doi.org/10.1007/s12250-020-00260-z

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