实验动物与比较医学2017，Vol.37Issue(5)：378-382,5.DOI:10.3969/j.issn.1674-5817.2017.05.007
小家鼠螺杆菌TaqMan-qPCR检测方法的建立
Development of TaqMan-qPCR for Detection of Helicobacter muridarum
摘要
Abstract
Objective To establish the sensitive and specific TaqMan-qPCR method for detection of Helicobacter muridarum.Method The specific primers were designed according to the Helicobacter muridarum gyrase B(GyrB) gene,specific fragment was amplified by PCR,and cloned into pMD-19T vector to construct the recombinant plasmid GyrB-19T,which was used as standard DNA of this qPCR method.The qPCR system was optimized and the sensitivity,specificity,repeatability and quantitative range of this method were evaluated.Result The quantitative standard curve from 2 × 108 copies/well to 2 × 102 copies/well of serial diluted plasmid DNAs showed that they had good liner correlation,the slope of the standard curve was-3.57,R2＞0.996,and the lowest detection limit reached 2 × 102 copies/well.Conclusion This qPCR method showed high sensitivity,specificity and stability,which will be utilized for qualitativeand quantitative detection of Helicobacter muridarum.关键词
小家鼠螺杆菌/TaqMan-qPCRKey words
Helicobacter muridarum/TaqMan-qPCR分类
农业科技引用本文复制引用
伍妙梨,朱余军,饶丹,袁文,王静,丛锋,黄韧,陈梅丽,郭鹏举..小家鼠螺杆菌TaqMan-qPCR检测方法的建立[J].实验动物与比较医学,2017,37(5):378-382,5.基金项目
广东省省级科技计划项目(2013B060400028 （）
2014A070705003 （）
2014B070706006) （）
