畜牧与兽医2025，Vol.57Issue(1)：74-78,5.
犬冠状病毒和犬细小病毒双重TaqMan荧光定量PCR检测方法的建立
Development of a duplex TaqMan real-time PCR assay for simultaneous detection and differentiation of canine coronavirus and canine parvovirus
宫英杰 1扈富哥 1王蕾 2毕振威 3钱晶 3王建发 4谭业平3
作者信息
- 1. 黑龙江八一农垦大学动物科技学院,黑龙江大庆 163319||江苏省农业科学院兽医研究所,江苏南京 210014
- 2. 南通伊仕生物技术股份有限公司,江苏南通 226010
- 3. 江苏省农业科学院兽医研究所,江苏南京 210014||兽用生物制品(泰州)国泰技术创新中心,江苏泰州 225300
- 4. 黑龙江八一农垦大学动物科技学院,黑龙江大庆 163319
- 折叠
摘要
Abstract
A dual TaqMan fluorescence quantitative PCR assay for detection of canine coronavirus and canine parvovirus was established in this study.Primers and probes were designed for 3'UTR of CCoV and VP2 of CPV.After the reaction system and conditions were optimized,the sensitivity,specificity and repeatability of the method were verified and clinical samples were tested.The results showed that the standard curves R2 of CCoV and CPV positive reference plasmids ranged from 1 ×107 copies/μL to 1×102 copies/μL,and the linear relationship was good.The minimum detection limits of CCoV and CPV were both 5 copies/μL.There was no cross-reaction with other pathogens,and the co-efficient of variation within and between groups was less than 1.38％.The results indicated that this method had high sensitivity,strong spe-cificity and good repeatability.Compared with ordinary PCR,the detection rate of CCoV and CPV using this method was 37.2％higher and 40.7％higher.Therefore,the PCR method established in this study had a good effect and could provide help for clinical diagnosis and epide-miological investigation.关键词
犬冠状病毒/犬细小病毒/双重实时荧光定量PCR/TaqMan探针Key words
canine coronavirus/canine parvovirus/dual real-time fluorescent quantitative PCR/TaqMan probe分类
畜牧业引用本文复制引用
宫英杰,扈富哥,王蕾,毕振威,钱晶,王建发,谭业平..犬冠状病毒和犬细小病毒双重TaqMan荧光定量PCR检测方法的建立[J].畜牧与兽医,2025,57(1):74-78,5.
