甲型H1N1流感病毒FAST-LAMP检测方法的建立.pdfVIP

生物技术通报 ·技术与方法· BIOTECHNOLOGY BULLETIN ( ) 2015, 31 5 :78-83 甲型 H1N1 流感病毒 FAST-LAMP 检测方法的建立 吕沁风  罗鹏  何蕾  杨永耀  郑伟  李莉  吴忠华  （浙江国际旅行卫生保健中心，杭州 310003） 摘 要 ： 建立一种新型等温扩增检测方法，用于提高等温扩增反应的检测速度，应用于甲型H1N1 流感病毒检测。根据HA 基因设计一组引物，包括外引物、 内引物、环引物及套内引物，套内引物的加入增加了引物与模板的接触位点，提高扩增效率， 缩短检测时间。结果显示，FAST-LAMP 检测法比普通LAMP 检测法时间缩短45 min 左右，比加入环引物的LAMP 检测方法缩短 2 拷贝。FAST-LAMP 是一种高效、更快的检测方法。 20 min，大大缩短了反应的检测时间。检测灵敏度可达到 1×10 关键词 ： FAST-LAMP ；甲型H1N1 流感病毒 ；检测 DOI ：10.13560/ki.biotech.bull.1985.2015.05.013 - The Establishment of FAST LAMP Method for Detecting Influenza A （ ） H1N1 Virus ü L Qinfeng Luo Peng He Lei Yang Yongyao Zheng Wei Li Li Wu Zhonghua （Zhejiang International Travel Healthcare Center ，Hangzhou 310003 ） Abstract: It was to establish a new loop-mediated isothermal amplification（FAST-LAMP）for rapidly detecting influenza A（H1N1） virus. A set of primers were designed according to HA gene, including the outer primers, internal primers, loop primers and nest internal primers. The nest internal primers increased the contact sites of primers and nucleic acid template, so that the amplification efficiency could be improved and detection time reduced. The FAST-LAMP method could be 45 less than LAMP method, and 20 minutes less than LAMP method with loop primers, i.e. detection time reduced significantly detecting sensitivity reached 1×102 copies cDNA template/tube. It proved that the established FAST-LAMP method is an efficient and reliable method for detecting influenza A（H1N1）virus. Key wor