小鼠肝炎病毒核酸快速检测方法的建立和应用  

Establishment and Application of Rapid Nucleic Acid Detection Methods for Mouse Hepatitis Virus

作　　者：熊炜[1] 蒋静[1] 张强[1] 刘俊平[1] 魏晓锋 黄忠荣[1] 李健[1] 胡建华 

机构地区：[1]上海出入境检验检疫局,上海200135 [2]上海实验动物研究中心,上海201203

出　　处：《实验动物科学》2013年第4期1-5,共5页Laboratory Animal Science

基　　金：上海市科委科研项目(No.11140901100和13DZ0502500)

摘　　要：为满足口岸对入境噬齿类动物快速检疫的需要,建立小鼠肝炎病毒(Mouse hepatitis virus,MHV)快速核酸检测方法。方法使用引物和TaqMan荧光探针设计软件,针对MHV保守基因设计引物和探针,建立了RT-PCR和Real-time RT-PCR方法检测噬齿类动物临床样本中MHV的方法。结果本研究建立的RT-PCR和Real-time RTPCR检测MHV方法具有良好的特异性和敏感性,通过将建立的方法应用于MHV阳性鼠临床样品的检测,证实两种MHV核酸检测方法具有良好的稳定性。结论本研究建立的RT-PCR和Real-time RT-PCR检测MHV的方法,适用于动物粪便、尿液等易于获得样本的检测。Objective To establish the methods of nucleic acid detection for mouse hepatitis virus （MHV） so as to meet the need of rapid quarantine for rodent animals in the port of entry.Method Using primer and TaqMan fluorescent probe software,the primers and probe were designed to target conserved gene of MHV.And methods of RT-PCR and Real-time RT-PCR using TaqMan probe were established to detect MHV from the clinical samples of rodent animals.Result The two established methods for MHV detection were proved to be specific with high sensitivity and efficiency.Using these two methods to detect clinical samples collected from MHV positive mouse,the results show RT-PCR and Real-time RT-PCR methods have high stability and reliablity.Conclusion The methods of RT-PCR and real-time RT-PCR to detect MHV were established,which are suitable to detect easily available samples such as feces and urine.

关 键 词：小鼠肝炎病毒 RT-PCR REAL-TIME RT-PCR 

分 类 号：Q95-33[生物学—动物学]