Subtyping of type A influenza by sequencing the variable regions of HA gene specifically amplified with RT-PCR
YAN An1, DING GuoHui2, ZHOU ZhenFeng1, DONG Hui1, ZHANG YaKun1, ZHU Lei3, HE YunGang3, ZHANG GuoQing4, LI YiXue2,4, SUN Bing2, HUANG Zhong2, LAN Ke2, JIN Li1,2,5, WANG HongYan5, WANG XiaoNing5,6, YANG Zhong4,5, ZHONG Yang4,5, DAI JianXin7,8, GUO YaJun7,8, WANG Hao7,8, CHE XiaoYan9, WU Fan10, YUAN ZhenGan10, ZHANG Xi10, CAO ZhiWei4,11, ZHOU XiaoNong12, ZHOU JiaHai13, MA ZhiYong14, TONG GuangZhi14, ZHAO GuoPing1,2,3,5, JIN WeiRong3 &|XIONG Hui1
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
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 pro-tein 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 de-tection 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.