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猪瘟病毒荧光定量RT-PCR检测方法的建立
引用本文:任夫波,张志,张丽丽,杨若松,韩艳,张燕霞,李晓成,单虎.猪瘟病毒荧光定量RT-PCR检测方法的建立[J].动物医学进展,2011,32(5):82-85.
作者姓名:任夫波  张志  张丽丽  杨若松  韩艳  张燕霞  李晓成  单虎
作者单位:任夫波,张丽丽,韩艳,REN Fu-bo,ZHANG Li-Li,HAN Yan(青岛农业大学,山东青岛,266109;中国动物卫生与流行病学中心,山东青岛,266032);张志,张燕霞,李晓成,ZHANG Zhi,ZHANG Yan-xia,LI Xiao-cheng(中国动物卫生与流行病学中心,山东青岛,266032);杨若松,YANG Ruo-song(中国动物卫生与流行病学中心,山东青岛,266032;西北农林科技大学,陕西杨凌,712100);单虎,SHAN Hu(青岛农业大学,山东青岛,266109)
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...

关 键 词:猪瘟病毒  荧光定量PCR  TaqMan探针

Establishmentt of Fluorescent Quantitative RT-PCR for Detection of Classical Swine Fever Virus
REN Fu-bo,ZHANG Zhi,ZHANG Li-Li,YANG Ruo-song,HAN Yan,ZHANG Yan-xia,LI Xiao-cheng,SHAN Hu.Establishmentt of Fluorescent Quantitative RT-PCR for Detection of Classical Swine Fever Virus[J].Progress In Veterinary Medicine,2011,32(5):82-85.
Authors:REN Fu-bo  ZHANG Zhi  ZHANG Li-Li  YANG Ruo-song  HAN Yan  ZHANG Yan-xia  LI Xiao-cheng  SHAN Hu
Institution:1(1.Qingdao Agricultural University,Qingdao,Shandong,266109,China;2.China Animal Health and Epidemiology Center,Qingdao,Shandong,266032,China;3.Northwest A & F University,Yangling,Shaanxi,712100,China)
Abstract: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.
Keywords:CSFV  fluorescent quantitative PCR  TaqMan probe
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