摘要
本研究根据BNoV和BKoV基因组的保守区域设计2对特异性引物,建立了能同时检测牛诺如病毒(Bovine norovirus,BNo V)和牛嵴病毒(Bovine kobuvirus,BKo V)的双重PCR检测方法。该方法能特异性扩增BNo V和BKo V的目的条带,且未扩增出其他犊牛腹泻病毒性病原;BNoV和BKoV的最低检测限分别为5.39×105 copies/μL和2.23×106 copies/μL;对临床采集的127份犊牛腹泻样品检测结果显示,BNoV的阳性率为6.30%(8/127),BKoV的阳性率为4.72%(6/127),两者与单一PCR检测结果相一致,符合率为100%。本研究建立的双重PCR方法具有良好的敏感性、特异性、重复性,可用于BNoV和BKoV的快速检测和流行病学调查。
To develop a PCR method for simultaneous detection of Bovine norovirus(BNoV)and Bovine kobuvirus(BKoV),two pairs of specific primers were designed based on the conserved regions of their genomes and amplified 542 bp fragment specific for BNoV and 216 bp fragment specific for BKoV.This PCR method was specific for BNoV and BKoV and no amplification for other calf diarrhea viruses.The sensitivity of this PCR method was determined to be 5.39×10~5 copies/μL for BNoV and 2.23×106 copies/μL for BKoV.Furthermore,127 clinical samples were collected from diarrheal calves and tested using this duplex PCR method.As a result,8 samples(6.30%,8/127)were positive for BNoV and 6 samples(4.72%,6/127)were positive for BKoV,which was consistent with single PCR results.Therefore,the duplex PCR method developed in this study had good sensitivity,specificity and repeatability and would be used for the rapid detection and epidemiological investigation of BNoV and BKoV.
作者
师志海
徐照学
兰亚莉
王文佳
SHI Zhi-hai;XU Zhao-xue;LAN Ya-li;WANG Wenjia(Institute of Animal Husbandry and Veterinary,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;College of Pharmaceutical Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation,Zhengzhou 450002,China)
出处
《中国动物传染病学报》
CAS
北大核心
2020年第1期28-32,共5页
Chinese Journal of Animal Infectious Diseases
基金
国家肉牛牦牛产业技术体系(CARS-37)
河南省农业科学院自主创新专项基金(2017ZC51,2018ZC56)
河南省“四优四化”科技支撑行动计划(2017-106-2130106)
