猪链球菌血清2型环介导等温扩增快速检测方法的建立  

Development of the loop-mediated isothermal amplification for rapid detection of Streptococcus suisserotype 2

作　　者：张九州[1] 李欢[1] 杨霞[1] 赵军[1] 陈陆[1] 常洪涛[1] 程海卫[1] 郑逢梅[1] 王川庆[1] 迪丽拜尔.阿木提 

机构地区：[1]河南农业大学,河南郑州450002 [2]阿克苏职业技术学院,新疆阿克苏843000

出　　处：《中国兽医科学》2012年第4期384-389,共6页Chinese Veterinary Science

基　　金：河南省科技攻关项目(0624030030);国家自然科学基金项目(30972187);国家农业科技成果转化基金项目(011GB2D00007)

摘　　要：为了建立快速、灵敏的猪链球菌血清2型(SS2)LAMP检测方法,根据GenBank登录的SS2特异的荚膜多糖(cps2 H)基因序列作为检测靶标,在其序列的保守区域设计LAMP引物,利用参考菌株S735基因组DNA为模板进行扩增,优化了LAMP的反应条件和反应体系,并进行了敏感性、特异性和重复性试验。结果,利用建立的LAMP方法对SS2进行扩增,扩增产物显色呈现阳性反应,电泳出现阶梯状条带,最低检测量为0.186fg/μL模板DNA,比常规PCR高1 000倍;且与其他常见的细菌无交叉反应。结果表明,建立的LAMP方法具有灵敏、特异、快速和重复性强等优点,适用于猪链球菌病的实验室快速检测。A loop-mediated isothermal amplification（LAMP） assay was developed for rapid,sensitive and specific detection of Streptococcus suis serotype 2（SS2）.A set of four primers were designed based on capsular polysaccharide（cps2 h）gene of SS2.LAMP was conducted under optimized conditions.Ladder-like DNA fragments were observed on agarose gel electrophoresis for amplified products from SS2 positive samples.The sensitivity of the developed LAMP was 1 000 times higher than that of the conventional PCR,with detection limit of 0.186 fg/μL.The whole detection process took only one hour.The LAMP described in this study was proved to be a sensitive,specific and rapid assay for the detection of SS2.It has the potential for both laboratorial and field detections of SS2.

关 键 词：猪链球菌血清2型 环介导等温扩增技术 快速检测 

分 类 号：S852.61[农业科学—基础兽医学]