猪瘟病毒荧光定量RT-PCR检测方法的建立  

Establishmentt of Fluorescent Quantitative RT-PCR for Detection of Classical Swine Fever Virus

作　　者：任夫波[1,2] 张志[2] 张丽丽[1,2] 杨若松[2,3] 韩艳[1,2] 张燕霞[2] 李晓成[2] 单虎[1] 

机构地区：[1]青岛农业大学,山东青岛266109 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]西北农林科技大学,陕西杨凌712100

出　　处：《动物医学进展》2011年第5期82-85,共4页Progress In Veterinary Medicine

摘　　要：本研究在CSFV 5′端非编码区设计一对引物和一条特异性TaqMan探针,以构建的重组质粒为标准品,建立了一种检测CSFV的荧光定量PCR方法（FQ-PCR）,对该方法进行特异性、敏感性和重复性试验。结果显示,可特异地检测CSFV,该方法在101~107拷贝/μL范围内具有良好的线性关系,灵敏度可达10拷贝/μL,重复检测的变异系数均小于5%。临床样品检测表明建立的方法与套式PCR灵敏度相近。结果表明该方法重复性良好,可用于临床诊断。A pair of primers and a specificity TaqMan probe were designed according to the published 5′ UTR sequences of CSFV and the recombingant plasmid was constructed as a standard control,then a fluorescent quantitaitive PCR method was developed,and tests of specificity,sensitivity and reproducibility were carried out.The results showed that the method could specifically detect CSFV,the sensitivity of this method was 10 copies/μL,the linear relation was excellent in a wide range of 101-107 copies/μL,and the coefficient of variation value was less than 5%.The sensitivity of FQ-RT-PCR is roughly equal to that of nPCR in detecting clinical sample.The results showed that the real time RT-PCR was repeatable,and could be used in clinical diagnosis.

关 键 词：猪瘟病毒 荧光定量PCR TAQMAN探针 

分 类 号：S852.651[农业科学—基础兽医学]