甲型H1N1流感病毒HA基因RT-LAMP检测方法的建立与初步应用  

Establish and Preliminary Application of RT-LAMP Technique in Detection for Influenza A(H1N1) HA Gene

作　　者：王翔[1] 张骞[1] 张钫[1] 袁武梅[1] 麻粉莲[1] 薛鹏浩[1] 赵智慧[1] 郑文芝[1] 郑丽舒[1] 

机构地区：[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,北京100052

出　　处：《生物技术通讯》2011年第5期696-699,708,共5页Letters in Biotechnology

基　　金：国家传染病防治科技重大专项(2009ZX10004-101;2008ZX10004-002)

摘　　要：目的:建立反转录环介导等温扩增技术(RT-LAMP)检测甲型H1N1流感病毒HA基因的方法,并用于检测临床样本。方法:通过在线软件Primer Explorer V4设计H1N1 HA基因的RT-LAMP引物,建立RT-LAMP检测方法,并评价其灵敏度和特异度。结果:与传统RT-PCR方法相比,RT-LAMP检测方法具备更高的灵敏度,达到10个拷贝,并且具有良好的特异性;在76份呼吸道感染儿童的咽拭子标本中检测到2份阳性,与RT-PCR方法检测结果相同。结论:建立的H1N1 RT-LAMP检测方法灵敏特异,简单快速,具备H1N1现场检测应用前景。Objective： To establish the methods of RT-LAMP assay in influenza virus A（H1N1） HA gene and use to detect clinical samples.Methods： RT-LAMP primer for HA of H1N1 was designed by using Primer Ex-plorer V4 on line,and the sensitivity and specificity of the established assay was evaluated.Results： Comparing with traditional RT-PCR,RT-LAMP showed fine specificity and higher sensitivity until 10 copy.By RT-LAMP as-say we got 2 positive samples in 76 clinical swabs from the children with respiratory infections.Conclusion： RT-LAMP assay for detection of H1N1 is sensitive,efficient,rapid and simple.These results imply potential on-the-spot detection of influenza H1N1 by RT-LAMP method.

关 键 词：甲型H1N1流感病毒 RT-PCR RT-LAMP 

分 类 号：Q78[生物学—分子生物学]