家兔须癣毛癣菌SYBR GreenⅠ荧光定量PCR检测方法的建立及初步应用  

Development and application of SYBR GreenⅠreal-time PCR for detecting Trichophyton mentagrophytes in rabbit

作　　者：刘月[1] 刘利强[1] 刘娜[1] 刘彦威[1] 刘建钗[1] 张永英[1] 朱美霞[1] 刘贵巧[1] 杜鹃[1] 

机构地区：[1]河北工程大学农学院动物医学重点实验室,河北邯郸056021

出　　处：《中国兽医科学》2015年第8期832-836,共5页Chinese Veterinary Science

基　　金：河北省自然科学基金项目(C2015402157);河北省教育厅重点项目(Z02014048);邯郸市科技局资助项目(1422101047-3)

摘　　要：为建立家兔须癣毛癣菌快速检测方法,根据GenBank上已发表的须癣毛癣菌rDNA内转录间隔区核苷酸序列设计了1对特异性引物,建立了检测须癣毛癣菌SYBR GreenⅠ实时荧光定量PCR方法,并用该方法对临床分离的须癣毛癣菌和病料进行检测。结果显示,该方法灵敏度高,对须癣毛癣菌的最低检出量为10copies/μL;特异性强,与犬小孢子菌、絮状表皮癣、红毛癣菌、白色念珠菌、烟曲霉等病原真菌没有交叉反应。该方法操作简单,耗时短,只需2h即可完成整个试验过程。提示该方法可用于须癣毛癣菌的检测。According to the ITS regions of rDNA of Trichophyton mentagrophytes in GenBank,a pair of special primers was designed. Then SYBR Green Ⅰ real-time PCR for detecting T. mentagrophytes in rabbit was established, the fragment of amplification should be 140 bp in length. The developed method was used to detect T. mentagrophytes strains and clinic samples. The result showed that SYBR Green Ⅰ PCR had many advantages such as rapidness, simple operation, high sensitivity and specificity, so it could be applied to clinic detection of T. mentagrophytes in rabbit.

关 键 词：家兔 须癣毛癣菌 SYBR GreenⅠ定量PCR 

分 类 号：S852.661[农业科学—基础兽医学]