摘要
为建立能同时检测牛诺如病毒(BNoV)和牛环曲病毒(BToV)的方法,根据BNoV和BToV基因组的保守区域设计2对特异性引物,预期特异性扩增BNoV和BToV的片段大小分别为542,698 bp,建立了BNoV和BToV的双重PCR检测方法。特异性检测结果显示,该方法能特异性扩增BNoV和BToV的目的条带,且未扩增出其他犊牛腹泻病原;对BNoV和BToV的最低检出量分别为1.90×10^4,1.86×10^4拷贝/μL。利用该方法对采自河南部分地区的184份犊牛腹泻样品的检测结果显示,BNoV和BToV的阳性检出率分别为11.41%和12.50%,与单一PCR检测结果一致。结果表明,本试验建立的方法对BNoV和BToV的鉴别诊断、混合感染和流行病学调查具有重要的应用价值。
To establish a PCR method for simultaneous detection of bovine norovirus(BNoV) and bovine torovirus(BToV),two pairs of specific primers were designed based on the conserved regions of BNoV and BToV genomes.The assay was highly specific to amplify the fragments of 542 bp for BNoV and 698 bp for BToV,respectively,and had no amplification for other related viruses.The sensitivity of the method was 1.90×10^4 copies/μL for BNoV and 1.89×10^4 copies/μL for BToV,as well as a good specificity.Furthermore,this duplex PCR method was used to test 184 clinical samples that collected from the diarrheic calves in Henan province.The positive rates of BNoV and BToV were 11.41% and 12.50%,respectively,which were consistent with single PCR results.Therefore,the duplex PCR method has a potential application in clinical diagnosis,differential diagnosis in mixed infections and epidemiological investigation of BNoV and BToV.
作者
王文佳
徐照学
张彬
孟红丽
王亚州
兰亚莉
师志海
WANG Wen-jia;XU Zhao-xue;ZHANG Bin;MENG Hong-li;WANG Ya-zhou;LAN Ya-li;SHI Zhi-hai(College of Pharmaceutical Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;Institute of Animal Husbandry and Veterinary,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation,Zhengzhou 450002,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2020年第5期892-896,共5页
Chinese Journal of Veterinary Science
基金
河南省重点研发与推广专项(科技攻关)资助项目(192102110074)
“十三五”国家重点研发计划资助项目(2018YFD051700)
广西水牛遗传繁育重点实验室开放课题资助项目(SNKF-2017-02)
国家现代农业肉牛牦牛产业技术体系资助项目(CARS-37)
河南省农业科学院自主创新专项基金资助项目(2017ZC51,2018ZC56)。
