中国动物传染病学报2017，Vol.25Issue(1)：19-25,7.
PEDV和TGEV双重TaqMan荧光定量一步法RT-PCR检测方法的建立及应用
DEVELOPMENT OF ONE STEP DUPLEX TAQMAN REAL-TIME PCR ASSAY FOR DETECTION OF PORCINE EPIDEMIC DIARRHEA VIRUS AND TRANSMISSIBLE GASTROENTERITIS VIRUS
摘要
Abstract
In the present study,one step duplex TaqMan fluorescent real-time RT-PCR assay of PEDV and TGEV was developed to detect and differentiate Porcine epidemic diarrhea virus,(PEDV) and Transmissible gastroenteritis virus,(TGEV).The specific primers and TaqMan fluorescent probes of these viruses were designed based on the bio-information analysis and then the amplification condition and the concentration of each virus were optimized.The result showed that the detection limits were 101 copies for both PEDV and TGEV and there was no cross reaction with other common viruses.Subsequently,one step Duplex TaqMan Real-time PCR assay was assembled into the PEDV and TGEV Detection Kit.The inter and intra-assay trials demonstrated that the variations were less than 0.58％ and 3.7％,respectively,indicating its good reproducibility.Total 97 samples were examined using this kit and the positive rate of PEDV and TGEV was 14.93％ higher than the current PCR method.These results demonstrated that this assay was a rapid,specific and sensitive method for detection of PEDV and TGEV.In conclusion,the method established in the present study can be used for rapid detection and epidemiology study of PEDV and TGEV.关键词
猪流行性腹泻病毒/猪传染性胃肠炎病毒/双重TaqMan实时荧光定量RT-PCRKey words
Porcine epidemic diarrhea virus/Transmissible gastroenteritis virus/duplex TaqMan real-time RT-PCR分类
农业科技引用本文复制引用
侯月娥,伍建敏,李中圣..PEDV和TGEV双重TaqMan荧光定量一步法RT-PCR检测方法的建立及应用[J].中国动物传染病学报,2017,25(1):19-25,7.基金项目
广东省科技计划项目——猪伪狂犬突变株的鉴定及新型、高效猪伪狂犬疫苗开发与产业化(2015B020203006) （2015B020203006）
