多基因表达系统在口岸流感病毒检测中的应用  

Study on the application of gene expression profiler genetic analysis system in detection of port influenza virus

作　　者：高国龙[1] 田绿波[1] 陈肖潇[1] 石莹[1] 樊学军[1] 杨雨[1] 

机构地区：[1]四川国际旅行卫生保健中心,四川成都610041

出　　处：《现代预防医学》2016年第16期3002-3007,共6页Modern Preventive Medicine

基　　金：国家质检总局科技计划项目(2014IK064);四川出入境检验检疫局科技计划项目(SK201402)

摘　　要：目的多基因表达分析系统(Gene expression Profiler genetic analysis system,Ge XP)是一种全新的基因表达分析平台,检测特异性强,检测速度和灵敏度明显提高。本研究利用Ge XP系统和多重PCR技术,结合我国口岸检疫查验主要呼吸道传染病现状,建立甲型流感病毒、乙型流感病毒、甲型H1N1病毒的检测技术方法。方法制备病毒标准品质粒,收集病原体样本,利用Gen Bank下载3种病毒的核苷酸序列,根据3种病毒的序列特征,利用Ge XP e Xpress Profiler设计特异性引物,优化条件使Ge XP系统能够特异性的检测对应病毒。结果优化后的引物能够检测3种病毒,产物经Ge XP检测,3个峰值位于166.04 bp、193.33 bp、251.42 bp,符合甲型流感病毒、乙型流感病毒、甲型H1N1病毒病原体质粒PCR产物条带大小,特异性强,无交叉反应。对已知临床样本的检测中,检出率高且特异性强。结论 Ge XP系统能够用于病原体高通量检测工作,有益于扩展检测途径,提高检测效率。Objective Gene expression profiler genetic analysis（Ge XP） using capillary electrophoresis separation technique is a new platform for gene expression analysis, with strong specificity,high detection speed and sensitivity. In this study, Ge XP technology and multiple PCR technology were used to establish the detection method of influenza A virus, influenza B virus,H1N1 virus combined with China＇s port quarantine inspection of major respiratory tract infectious diseases. Methods The nucleotide sequence of the virus was downloaded using Gen Bank. Preparation of viral standard plasmid and collection of pathogen samples were performed. According to the sequence characteristics of three kinds of viruses and designed specific primers using Ge XP e Xpres, conditions were optimized to detect the corresponding virus in Ge XP system. Results The optimized primers could detect the virus with strong specificity and no cross reaction,three peaks in 166.04 bp, 193.33 bp,251.42 bp were detected by Ge XP, in line with virus plasmid PCR product band size of influenza A virus, influenza B virus,H1N1 virus. At the same time, detection rate and the specificity were strong in the detection of clinical samples. Conclusion Ge XP system can be used for high throughput detection of pathogens,which was beneficial to extend the detection method and improve the detection efficiency.

关 键 词：输入性传染病 流感病毒 多基因表达分析系统 检测方法 

分 类 号：R183.3[医药卫生—流行病学]