牛传染性鼻气管炎gE基因TaqMan-MGB与SYBR Green Ⅰ实时荧光定量PCR方法的建立及应用  

Establishment and application of TaqMan-MGB and SYBR GreenⅠreal-time fluorescent quantitative PCR methods for the gE gene of infectious bovine rhinotracheitis

作　　者：袁浩芳 朱晓晖 张雨 李焱 李可伟 胡剑武 李家奎[1] YUAN Haofang;ZHU Xiaohui;ZHANG Yu;LI Yan;LI Kewei;HU Jianwu;LI Jiakui(School of Animal Science and Technology,School of Animal Medicine,Huazhong Agricultural University,Wuhan 430070,China;Veterinary Service Bureau of Tibet Autonomous Region,Lhasa 850000,China)

机构地区：[1]华中农业大学动物科学技术学院动物医学院,湖北武汉430070 [2]西藏自治区农业农村厅兽医局,西藏拉萨850000

出　　处：《畜牧与兽医》2023年第11期110-116,共7页Animal Husbandry & Veterinary Medicine

基　　金：西藏自治区科技厅重点项目(XZ202101ZD0002N);西藏自治区重点研发计划(XZ202001ZY0044N);国家肉牛牦牛产业技术体系项目(CARS-37);西藏自治区科技厅2023年度重点研发计划项目(XZ202301ZY0016N)。

摘　　要：为建立检测牛传染性鼻气管炎病毒(IBRV)的荧光定量检测方法,本研究根据IBRV gE基因的序列设计了相应的探针和引物,分别建立TaqMan-MGB探针和SYBR GreenⅠ荧光定量PCR方法,并对2种方法进行了综合比较。结果显示,TaqMan-MGB探针和SYBR GreenⅠ定量荧光PCR方法的灵敏度分别达到1.0×10^(1) copies/μL和1.0×10^(2) copies/μL;2种荧光定量方法对除IBRV的其他牛病毒和细菌检测结果均为阴性;重复性检测中变异系数均小于2.3%;检测了130份来自西藏的牦牛样品,2种荧光定量PCR阳性检出率均为100%。以上结果表明,2种荧光定量方法都可用于检测IBRV,且TaqMan-MGB荧光定量PCR敏感性高于SYBR GreenⅠ荧光定量PCR。In order to establish a fluorescence quantification method for detection of infectious bovine rhinotracheitis virus(IBRV),we de⁃signed probes and primers based on the sequence of the IBRV gE gene,established TaqMan probe and SYBR GreenⅠfluorescence quantifi⁃cation PCR methods respectively,and compared the sensitivity,specificity and reproducibility of the two methods.The results showed that the sensitivity of the TaqMan probe and SYBR GreenⅠfluorescence PCR methods reached 1.0×101 copies/μL and 1.0×102 copies/μL,re⁃spectively.The results of the two fluorescence quantification methods were negative for all bovine viruses and bacteria except IBRV.Their co⁃efficients of variation were less than 2.3%in the reproducibility tests.130 suspected clinical materials were tested,and the positive detection rate of both the fluorescent PCR methods was 100%.The present results indicated that both fluorescence methods could be used to detect IBRV,and the TaqMan quantitative fluorescence PCR method was more sensitive than the SYBR GreenⅠquantitative fluorescence PCR method.

关 键 词：牛传染性鼻气管炎病毒 TaqMan-MGB荧光定量PCR SYBR GreenⅠ荧光定量PCR 

分 类 号：S858[农业科学—临床兽医学]