中国动物检疫2024，Vol.41Issue(4)：91-95,5.DOI:10.3969/j.issn.1005-944X.2024.04.017
猫冠状病毒和猫细小病毒双重荧光定量PCR方法的建立
Development of a Duplex Fluorescent Quantitative PCR Assay for Feline Coronavirus and Feline Parvovirus
郭存杰 1王蕾 2毕振威 3钱晶 3张传美 4谭业平3
作者信息
- 1. 青岛农业大学动物医学院,山东青岛 266109||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
- 4. 青岛农业大学动物医学院,山东青岛 266109
- 折叠
摘要
Abstract
In order to develop a duplex fluorescent quantitative PCR assay to detect feline coronavirus(FCoV)and feline parvovirus(FPV),the conservative regions of FCoV 3'UTR and FPV VP2 gene were selected to design two pairs of specific primers and TaqMan-MGB probes,respectively,followed by optimization of the reaction system and conditions.Then the sensitivity,specificity and repeatability were tested to explore the feasibility of the established assay.The results revealed that FCoV and FPV could be specifically detected by the assay,while no cross-reaction with feline herpesvirus,feline calici virus,feline rotavirus,Mycoplasma,Chlamydia,Bordetella,canine adenovirus and canine parainfluenza virus,was found.The detection limits for both FCoV and FPV were 1 copies/μL;the variable coefficients of inter-group and intra-group repeated tests of FCoV and FPV positive reference plasmids were both less than 3%;and it was found that,through detection of 35 clinical samples,the positive rate detected by the established method was higher than that by the traditional PCR.In conclusion,the established method,with the advantages of high sensitivity,good specificity and stability,could be used for early differential diagnosis of clinical infection with FCoV and FPV.关键词
猫冠状病毒/猫细小病毒/双重荧光定量PCRKey words
FCoV/FPV/duplex fluorescent quantitative PCR分类
农业科技引用本文复制引用
郭存杰,王蕾,毕振威,钱晶,张传美,谭业平..猫冠状病毒和猫细小病毒双重荧光定量PCR方法的建立[J].中国动物检疫,2024,41(4):91-95,5.
