裂谷热病毒两种PCR检测方法的建立及比较分析  

The comparation of two PCR methods for detection of rift valley fever virus

作　　者：冷依伊 王印[1,2] 项明源 张鹏飞[1,2] 蒋子睿 江地科 杨泽晓[1] 姚学萍[1] LENG Yi-yi;WANG Yin;XIANG Ming-yuan;ZHNAG Peng-fei;JIANG Zi-rui;JIANG Di-ke;YANG Ze-xiao;YAO Xue-ping(College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China)

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《中国预防兽医学报》2018年第6期504-508,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"十二五"国家科技支撑计划(2013BAD12B04)

摘　　要：裂谷热(RVF)具有较高的发病率与死亡率,极大危害着当地养殖业和生态系统的稳定。为建立一种能够快速有效检测裂谷热病毒(RVFV)的方法,本研究根据RVFV的保守序列设计了特异性引物与探针,分别建立了探针PCR和荧光PCR,并对这两种检测方法进行比较分析。结果显示,两种检测方法均不存在非特异扩增;最低检测浓度分别为1.03×10^(-2)拷贝/μL和1.03×10~0拷贝/μL,前者比后者有更高的灵敏度;探针PCR和荧光PCR组内和组间变异系数均小于3%。实验表明,探针PCR具有特异性强、灵敏度高、稳定性好、准确度高、检测速度快等优点。因此,探针PCR是一种灵敏度和可靠性均较高的RVFV检测方法。Rift Valley fever （RVF） has high morbidity and mortality, which seriously endangers the stability of local livestock and ecosystem. In order to establish a rapid and effective method for detection of RVF virus （RVFV）, the TaqMan real-time PCR, fluorescence SYBR Green I based quantitative PCR （FQ PCR） were established with the specific primers and probes designed according to the conserved sequence of RVFV, and compared for the RVFV detections. The results showed that TaqMan real-time PCR had the advantages of strong specificity, high sensitivity and stability with fast detection speed. The results of this study showed that although the TaqMan real-time PCR and FQ PCR were less than 3% for the coefficient of variation within and between groups, but the former had higher sensitivity than the latter, and the minimum detectable concentration of the two detection methods were 1.03×10^-2 copies/μL and 1.03 ×10^0 copies/μL of RVFV for the TaqMan real-time PCR and FQ PCR, and there was no nonspecific amplification. Therefore, the TaqMan real-time PCR was a high priority method for RVFV detection.

关 键 词：裂谷热病毒 探针PCR 荧光PCR 

分 类 号：S852.65[农业科学—基础兽医学]