荧光探针PCR检测牛嵴病毒的方法建立  

Establishment of a TaqMan PCR assay for detecting bovine kobuvirus

作　　者：师志海[1] 王文佳[2] 孟红丽[1] 王亚州[1] 闫祥洲[1] 邱祥琦 程燕 滑留帅[1] 施巧婷[1] 徐照学[1] SHI Zhihai;WANG Wenjia;MENG Hongli;WANG Yazhou;YAN Xiangzhou;QIU Xiangqi;CHENG Yan;HUA Liushuai;SHI Qiaoting;XU Zhaoxue(Institute of Animal Husbandry and Veterinary Medicine/Henan Key Laboratory of Farm Animal,Henan Academy of Agricultural Sciences Breeding and Nutritional Regulation,Zhengzhou Key Laboratory of Veterinary Bioinformatics,Zhengzhou 450002,China;College of Veterinary Medicine and Pharmaceutical Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China)

机构地区：[1]河南省农业科学院畜牧兽医研究所河南省畜禽繁育与营养调控重点实验室/郑州市兽医生物信息重点实验室,河南郑州450002 [2]河南牧业经济学院动物医药学院,河南郑州450046

出　　处：《中国兽医学报》2022年第4期657-661,共5页Chinese Journal of Veterinary Science

基　　金：国家肉牛牦牛产业技术体系基金资助项目(CARS-37);国家重点研发计划基金资助项目(2018YFD051700);河南省重点研发与推广专项基金资助项目(212102110360)。

摘　　要：牛嵴病毒(bovine kobuvirus,BKoV)是与犊牛腹泻密切相关的病原,试验针对BKoV 3D基因设计特异性引物和探针,建立快速检测BKoV的实验室方法。该方法仅能特异性扩增BKoV,对其他犊牛腹泻病原无扩增,特异性较强;对BKoV质粒标准品的最低检出限为11.0拷贝/μL,灵敏性较强;且组内与组间变异系数均小于2%,重复性较好。对采集自河南省的221份犊牛腹泻临床样本进行检测,BKoV的核酸检出率为9.05%(20/221)。本研究建立的检测方法为BKoV的临床快速检测和流行病学调查提供了有效手段。Bovine kobuvirus(BKoV)is an agent closely related to calf diarrhea.Specific primers and probe were designed based on the sequence of 3 D gene of BKoV,and a TaqMan fluorescent quantitative PCR method for rapid detection of BKoV was established in this study.The results showed that only BKoV was specifically amplified,no amplification curve for other calf diarrhea pathogens,strong specificity;the detection minimum limit of BKoV was 11.0 copies/μL,high sensitivity;the coefficient of variation within and between groups were less than 2%,high repeatability.221 fecal samples collected from the diarrheic calves in Henan Province were detected,and the detection rate of BKoV was 9.05%(20/221).The detection method established in this study provides an effective means for the rapid clinical detection and epidemiological investigation of BKoV infections.

关 键 词：牛嵴病毒 快速检测 荧光探针 

分 类 号：S852.65[农业科学—基础兽医学]