2006年、2007年广州大学城流感病毒的分子生物学检测及基因分析──附83份咽拭子标本检测报告  

Molecular biological detection and genetic analysis of influenza virus in 2006 and 2007 in Guangzhou Universty City

作　　者：吴新伟[1] 伍业健[1] 陈艺韵[1] 蒋力云[1] 李铁钢[1] 何丽娟[1] 王玉林[1] 

机构地区：[1]广州市疾病预防控制中心,510080

出　　处：《新医学》2009年第7期433-435,454,共4页Journal of New Medicine

基　　金：广东省科技计划资助项目(2008B030303041);广州市科技计划资助项目(2006J1-C0141;广州市医学科学技术研究重点资助项目(2006-ZDi-10)

摘　　要：目的:应用分子生物学方法了解2006年与2007年引发广州大学城流行性感冒(流感)疫情的病原体。方法:在3个校区采集流感样症状患者的咽拭子标本,使用A、B型流感病毒荧光PCR检测试剂盒,按照试剂盒说明书进行检测,以Ct值小于30判定为阳性,初步鉴别为流感病毒后,再用荧光PCR法进行病毒型别鉴定,并与MDCK细胞培养法及红细胞凝集抑制试验进行比较。同时用逆转录PCR法对流感病毒血凝素(HA)基因进行扩增,并测序分析其同源性。结果:83份咽拭子标本中,MDCK细胞培养法的检测阳性率为47%,而荧光PCR法的检测阳性率为58%。引起2006年和2007年广州大学城流感爆发疫情的分别是H1和H3亚型毒株。2006年与2007年广州大学城不同校区的流感病毒株HA基因的同源性分别为96.4%及99.2%～99.6%。结论:2006年和2007年广州大学城爆发流感疫情的病原体分别为H1和H3亚型流感病毒,同一年内发生在大学城不同校区的流感疫情是由同一病毒株引起的。荧光PCR法检测需时短,特异度和敏感度较高,不仅能快速准确地检测流感病毒,还能针对不同亚型进行分型。Objective： To study the pathogens of influenza outbreaks in year 2006 and 2007 in Guang- zhou University City by molecular biological methods. Methods： Throat swabs from influenza-like symptom of pa- tients were collected in 3 campuses. The influenza viruses were first identified by A or B type fluorescent PCR kits, under the the introduction of the kits and positive standard as Ct less than 30, then types of virus were identified by fluorescent PCR, and the results were compared with that of MDCK cell culture method and hemagglutination inhi- bition test. The hemagglutinin gene of influenza virus was amplified by RT-PCR, and sequenced and homological rate was analyzed. Results ： Eighty-three throat swabs were examined, the positive rate of MDCK cells culture was 47%, and that of fluorescent PCR was 58%. The influenza virus strains caused the outbreaks in Guangzhou Uni- versity City in 2006 and 2007 were identified as H1 subtype and H3 subtype, respectively. The homological rate of hemagglutinin gene of influenza viruses from different campuses was 96. 4% in 2006, and 99.2% ~ 99.6% in 2007. Conclusion： The pathogens of influenza outbreaked in 2006 and 2007 in Guangzhou University City were H1 subtype and H3 subtype influenza viruses, respectively. The outbreak of influenza in different campuses of Guang- zhou University City in same year was caused by one same influenza virus strain. Fluorescent PCR method, with short operation time and high specificity and sensibility, can be used not only in the quick detection, but also in the subtyping of influenza virus.

关 键 词：流行性感冒 流行性感冒病毒 病毒分离 荧光聚合酶链反应 红细胞凝集抑制试验 基因测序 

分 类 号：R450[医药卫生—治疗学]