犬细小病毒昆明流行株的分离鉴定与VP2基因序列分析

doi:10.16431/j.cnki.1671-7236.2019.01.024

Abstract

Abstract:

In order to understand the prevalence and antigenic variation of canine parvovirus (CPV) in Kunming,and select and develop vaccines pertinently,six suspected CPV feces were collected from some pet hospitals in Kunming for virus isolation and culture in this study.The obtained suspected virus solution was subjected to PCR,IFA,hemagglutination titer measurement,TCID₅₀ measurement,and VP2 gene sequence analysis.The results showed that the isolated virus could produce obvious cytopathic effect (CPE) in F81 cat kidney cells.The length of PCR product was 164 bp.The F81 cat kidney cells infected by IFA showed green fluorescence and the blood coagulation titer was 1:512.The TCID₅₀ of the virus was 10^-4.375/0.1 mL,and the length of VP2 gene was 1 755 bp.The VP2 structural protein of the isolate was sequenced and analyzed,and the isolate was determined to be CPV-2a subtype,and there were five variant amino acid sites compared with the standard strain,which belonged to the mutant strain.The sequencing results were compared with the commercial vaccine strains and the domestic reference strains.The results showed that the nucleotide homology with the attenuated vaccine Intervet/vaccine/06 and Pfizer/vaccine/06 were both 98.7%;The sequence homology was 98.2% to 99.5%,which had the highest homology with ANTU-1 and WH02/06 strains.It was closely related to ANTU-1 strain,WH02/06 strain and BJ01 strain,and was located in the same cluster of evolutionary tree.This study was of great significance for monitoring the genetic variation trend of CPV and the development of vaccines.

Key words: canine parvovirus (CPV); isolation; identification; VP2 gene

CLC Number:

S852.65⁺5

ZHAO Yiqin, ZENG Mengying, GUO Juan, ZHANG Di, GAO Hong, YAN Yulin. Isolation and Identification of Kunming Popular Strain of Canine Parvovirus and Sequence Analysis of VP2 Gene[J]. , 2019, 46(1): 206-214.

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Isolation and Identification of Kunming Popular Strain of Canine Parvovirus and Sequence Analysis of VP2 Gene

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