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首页> 中文期刊> 《农业科学学报（英文版）》 >Nested RT-PCR method for the detection of European avian-like H1 swine inlfuenza A virus

Nested RT-PCR method for the detection of European avian-like H1 swine inlfuenza A virus

         
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摘要

Swine inlfuenza A virus (swine IAV) circulates worldwide in pigs and poses a serious public health threat, as evidenced by the 2009 H1N1 inlfuenza pandemic. Among multiple subtypes/lineages of swine inlfuenza A viruses, European avian-like (EA) H1N1 swine IAV has been dominant since 2005 in China and caused infections in humans in 2010. Highly sensitive and speciifc methods of detection are required to differentiate EA H1N1 swine IAVs from viruses belonging to other lineages and subtypes. In this study, a nested reverse transcription (RT)-PCR assay was developed to detect EA H1 swine IAVs. Two primer sets (outer and inner) were designed speciifcaly to target the viral hemagglutinin genes. Speciifc PCR products were obtained from al tested EA H1N1 swine IAV isolates, but not from other lineages of H1 swine IAVs, other subtypes of swine IAVs, or other infectious swine viruses. The sensitivity of the nested RT-PCR was improved to 1 plaque forming unit (PFU) mL–1which was over 104 PFU mL–1 for a previously established multiplex RT-PCR method. The nested RT-PCR results obtained from screening 365 clinical samples were consistent with those obtained using conventional virus isolation methods combined with sequencing. Thus, the nested RT-PCR assay reported herein is more sensitive and suitable for the diagnosis of clinical infections and surveilance of EA H1 swine IAVs in pigs and humans.

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  • 来源
    《农业科学学报（英文版）》 |2016年第5期|1095-1102|共8页
  • 作者

    WEI Yan-di; PEI Xing-yao; ZHANG Yuan; YU Chen-fang; SUN Hong-lei; LIU Jin-hua; PU Juan;

  • 作者单位

    Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture/State Key Laboratory of Agrobiotechnology/Colege of Veterinary Medicine, China Agricultural University, Beijing 100193, P.R.China;

    Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture/State Key Laboratory of Agrobiotechnology/Colege of Veterinary Medicine, China Agricultural University, Beijing 100193, P.R.China;

    ChinaInstitute of Veterinary Drug Control, Beijing 100086, P.R.China;

    Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture/State Key Laboratory of Agrobiotechnology/Colege of Veterinary Medicine, China Agricultural University, Beijing 100193, P.R.China;

    Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture/State Key Laboratory of Agrobiotechnology/Colege of Veterinary Medicine, China Agricultural University, Beijing 100193, P.R.China;

    Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture/State Key Laboratory of Agrobiotechnology/Colege of Veterinary Medicine, China Agricultural University, Beijing 100193, P.R.China;

    Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture/State Key Laboratory of Agrobiotechnology/Colege of Veterinary Medicine, China Agricultural University, Beijing 100193, P.R.China;

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