犬细小病毒CPV-2c型亚洲株分离鉴定及VP2基因序列分析
Identification and VP2 Gene Sequence Analysis of Canine Parvovirus CPV-2c Asian Strain Isolation
李昀真 1李双双 1丛培强 2曹海旭 1鲁荣光 1廉士珍 1张海玲 1李虹晔 1胡博 1白雪1
作者信息
- 1. 中国农业科学院特产研究所,农业农村部经济动物疫病重点实验室,吉林 长春 130112
- 2. 山东省威海市文登区畜牧兽医技术服务中心,山东 威海 264400
- 折叠
摘要
为探明北京某犬场犬的死亡原因,从患有肠道出血的犬肛拭子中分离出1株犬细小病毒,试验通过PCR、血凝和间接免疫荧光试验方法对毒株进行鉴定,并对VP2基因进行克隆测序和序列分析,确定其遗传分支.结果表明,该分离株属于犬细小病毒CPV-2c型,命名为CPV-BJ21株.应用犬细小病毒单克隆抗体进行间接免疫荧光检测,结果为阳性;基因序列分析表明,分离株与犬细小病毒为同一进化分支,VP2基因核苷酸序列与中国四川的CPV-2c(MH476581.1)同源性达99％,与亚洲分离的犬细小病毒之间亲缘关系较近.VP2氨基酸序列在第A5G、S297A、D426E位出现了氨基酸突变;在F81细胞上传3代后,病毒液的血凝效价稳定于210.本研究可为犬细小病毒的流行情况及新疫苗的研究提供参考依据.
Abstract
Absrtact:To investigate the unknown death of dogs in Beijing kennel.A strain of CPV was isolated from the anal swabs of dogs which suf-fering from intestinal bleeding in a Beijing dog farm.The strain was identified by PCR,hemagglutination and indirect immunofluorescence.The VP2 gene were amplified and cloned by PCR,determined its genetic branch by sequence analyse.The results showed that the isolate strain belonged to Canine parvovirus type 2c and was named CPV-BJ21 strain.The indirect immunofluorescence results of the canine mono-clonal antibody were positive.Gene sequence analysis showed that CPV-BJ21 and CPV were the same evolutionary branch,the nucleotide sequence of VP2 gene had 99％homology with cpv-2c(MH476581)in Sichuan,China;CPV-BJ21 with close genetic relationship with Asian strains.There were amino acid mutations at A5G,S297A and D426E in VP2 amino acid sequence.After 3 generations of superculture of F81 cells,the hemagglutination titer of virus solution was stable at 210.This study could provide a reference for the prevalence of CPV and the development of new vaccines.
关键词
犬细小病毒/分离鉴定/VP2基因/遗传进化分析Key words
canine parvovirus/isolation and identification/VP2 gene/phylogenetic analysis引用本文复制引用
基金项目
国家自然科学基金(32100408)
吉林省科技发展计划(20210509057RQ)
农业部动物病原生物学重点实验室开放基金(BYSWX 2021 KFKT01)
出版年
2024
国家科技期刊平台
NSTL
万方数据