聂凯;王大燕;秦萌;高荣保;王淼;邹淑梅;韩峰;赵翔;李希妍;舒跃龙;马学军;

• 1:中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室

摘要(Abstract)：

建立了一种基于颜色判定的简单、快速和灵敏的检测方法,即环介导逆转录等温核酸扩增技术(RT-LAMP)应用于人甲型H1N1流感病毒基因检测。该技术使用对应于人甲型H1N1流感病毒HA序列中8个基因区段的6条特异引物,在等温条件下(65℃)进行核酸扩增反应1.5h,在扩增前加入染料HNB(羟基萘酚蓝)作为反应指示剂,以HNB的颜色变化做为结果判定标准并经琼脂糖凝胶电泳验证。文中利用这种技术对不同来源及亚型的流感病毒进行了特异性分析,对体外转录的人甲型H1N1流感病毒HA基因RNA的系列稀释物进行了灵敏度分析,成功检测美国CDC提供的人甲型H1N1流感病毒标准品,利用RT-LAMP和RT-PCR同时检测了30份人甲型H1N1和26份季节性流感咽拭子标本。结果显示RT-LAMP方法特异性高,灵敏度可达到60个拷贝RNA分子水平,对临床标本的检出率与常规RT-PCR法相当,利用650nm的比色分析通过标准曲线可以实现对样品的定量。因此,基于颜色判定的环介导逆转录等温扩增方法可用于人甲型H1N1流感病毒感染的快速筛选,具有在基层疾病预防控制中心流感监测网络实验室和哨点医院推广和应用的潜力。

关键词(KeyWords)： 等温扩增;人甲型H1N1流感病毒

Abstract:

Keywords:

基金项目(Foundation): 传染病防治重大专项(2009ZX10004-101;2008ZX10004-002;2008ZX10004-001;2008ZX10004-004)

作者(Authors): 聂凯;王大燕;秦萌;高荣保;王淼;邹淑梅;韩峰;赵翔;李希妍;舒跃龙;马学军;

DOI: 10.13242/j.cnki.bingduxuebao.002068

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