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Subtyping of type A influenza by sequencing the variable regions of HA gene specifically amplified with RT-PCR
Authors:An Yan  GuoHui Ding  ZhenFeng Zhou  Hui Dong  YaKun Zhang  Lei Zhu  YunGang He  GuoQing Zhang  YiXue Li  Bing Sun  Zhong Huang  Ke Lan  Li Jin  HongYan Wang  XiaoNing Wang  Zhong Yang  Yang Zhong  JianXin Dai  YaJun Guo  Hao Wang  XiaoYan Che  Fan Wu  ZhenGan Yuan  Xi Zhang  ZhiWei Cao  XiaoNong Zhou  JiaHai Zhou  ZhiYong Ma  GuangZhi Tong  GuoPing Zhao  WeiRong Jin  Hui Xiong
Institution:1. Shanghai-MOST Key Laboratory for Disease and Health Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, 201203, China
2. Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences (Lab of Computational Genomics, CAS-MPG Partner Institute of Computational Biology Center of Bioinformatics, Key Lab of System Biology; Lab of Synthetic Biology, Institute of Plant Physiology & Ecology; Institute Pasteur), Shanghai, 200031, China
3. National Engineering Research Center for Biochip Technology, Shanghai Biochip Co. Ltd, Shanghai, 201203, China
4. Shanghai Center for Bioinformation Technology, Shanghai, 200235, China
5. School of Life Sciences, Fudan University, Shanghai, 200433, China
6. School of Life Sciences and Technology, South China University of Technology, Guangzhou, 510641, China
7. Cancer Institute, Second Military Medical University, Shanghai, 200433, China
8. National Engineering Research Center for Antibody Drugs, Shanghai, 201203, China
9. Zhujiang Hospical, Southern Medical University, Guangzhou, 510282, China
10. Shanghai Municipal Center for Disease Control & Prevention, Shanghai, 200336, China
11. School of Life Sciences and Technology, Tongji University, Shanghai, 200092, China
12. National Institute for Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai, 200025, China
13. Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, 200032, China
14. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China
Abstract:The outbreak of a novel influenza A (H1N1) virus across the globe poses a threat to human health. It is of paramount importance to develop a rapid, reliable and inexpensive diagnostic procedure. Based on the bioinformatic information from public database, primers specific for influenza A virus surface protein haemagglutinin (HA) of several subtypes (including H1, H2, H3, H5, H7 and H9) were designed. Primer-specific PCR products were subjected to sequencing for accurately distinguishing H1 and H3 subtypes from others. This sequencing-based detection method will not only be applied to rapid detection and simultaneous subtype identification of new influenza A virus H1N1, but also provide the strategies to monitor other new types of influenza virus with explosive potential. Contribute equally to this work.
Keywords:influenza A virus            HA gene  H1 and H3 subtypes  specific amplification by RT-PCT  DNA sequencing
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