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.
作者
冷依伊
王印
项明源
张鹏飞
蒋子睿
江地科
杨泽晓
姚学萍
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)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
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)
关键词
裂谷热病毒
探针PCR
荧光PCR
Rift Valley fever virus
TaqMan probe real-time PCR
fluorescence SYBR Green I based quantitative PCR
作者简介
冷依伊（1993-），女，四川成都人，硕士研究生，主要从事动物传染病病原分子生物学研究．;通信作者：E-mail：18227553251@163．com
