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牛传染性鼻气管炎病毒实时荧光定量PCR检测方法的建立
引用本文:唐泰山,邓碧华,王凯民,张常印,雷治海.牛传染性鼻气管炎病毒实时荧光定量PCR检测方法的建立[J].动物医学进展,2009,30(4).
作者姓名:唐泰山  邓碧华  王凯民  张常印  雷治海
作者单位:1. 江苏出入境检验检疫局动物检疫实验室,江苏南京,210001
2. 江苏出入境检验检疫局动物检疫实验室,江苏南京,210001;南京农业大学动物医学院,江苏南京,210095
3. 南京农业大学动物医学院,江苏南京,210095
基金项目:江苏检验检疫局科技项目 
摘    要:根据牛传染性鼻气管炎病毒(IBRV)保守基因gB和gE序列,分别设计两对引物及其相应的TaqMan探针,建立、优化反应体系后,利用10倍稀释的病毒进行扩增以检测其灵敏度,同时对伪狂犬病病毒(PRV)、马立克病病毒(MDV)、鸭瘟病毒(DPV)进行特异性检测.结果显示,建立的扩增gB和gE基因的荧光PCR可用于检测IBRV,其灵敏度为0.02 TCID50,而与PRV等非IBRV无交叉反应.本研究所建立的荧光PCR具有快速、灵敏、准确、低污染等优点,可用于IBRV的检测.

关 键 词:牛传染性鼻气管炎病毒  实时荧光定量PCR  特异性  灵敏度

Establishment of Real-time RT-PCR Assays for Detecting Bovine infectious rhinotracheitis virus
TANG Tai-shan,DENG Bi-hua,WANG Kai-min,ZHANG Chang-yin,LEI Zhi-hai.Establishment of Real-time RT-PCR Assays for Detecting Bovine infectious rhinotracheitis virus[J].Progress In Veterinary Medicine,2009,30(4).
Authors:TANG Tai-shan  DENG Bi-hua  WANG Kai-min  ZHANG Chang-yin  LEI Zhi-hai
Institution:1.Lab of Animal Quarantine;APHIC;Jiangsu Entry-Exit Inspection And Quarantine Bureau;Nanjing;Jiangsu;210001;China;2.College of Veterinary Medicine;Nanjing Agricultural University;210095;China
Abstract:Two pairs of real-time RT-PCR primers and probes,gB and gE were designed and synthesized based on the nucleotide sequences of infectious bovine rhinotracheitis virus.The minimal detection threshold of gB and gE real-time PCR were approximately 0.02 TCID50.The specificity of the two methods was demonstrated by detecting pseudorabies virus(PRV),Marek's disease virus(MDV) and duck plague virus(DPV).It was concluded that the real time RT-PCR could be an effective method for rapid detection of IBRV.
Keywords:Infectious bovine rhinotracheitis virus  real-time PCR  specificity  sensitivity  
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