Development of a Duplex Fluorescent Quantitative PCR Assay for Feline Coronavirus and Feline Parvovirus
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.
国家科技期刊平台
NSTL
万方数据
维普
