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猪A群轮状病毒的分离与鉴定
引用本文:库旭钢,张坤,刘羽茜,何启盖.猪A群轮状病毒的分离与鉴定[J].畜牧兽医学报,2012,43(2):275-281.
作者姓名:库旭钢  张坤  刘羽茜  何启盖
作者单位:农业微生物学国家重点实验室,武汉430070;华中农业大学动物医学院,武汉430070
基金项目:国家生猪现代产业技术体系,广东省中国科学院全面战略合作项目
摘    要:目前国内对猪轮状病毒的研究较少,作者旨在对猪A群轮状病毒进行分离与鉴定,为后续猪轮状病毒致病机理和分子生物学特性研究奠定基础.用胰蛋白酶处理RT-PCR检测猪A群轮状病毒病原阳性的临床腹泻粪样,然后接种长成单层的MA-104细胞,进行病毒分离传代.再对分离毒株进行常规RT-PCR鉴定和电镜观察,并对分离毒株的VP6、VP7和VP8基因进行测序及序列分析.结果成功分离猪A群轮状病毒TM-a株,经序列同源性分析发现,与TM-a株VP6、VP7和VP8基因同源性最高的毒株分别为中国北京人源LL3354株、印度人源RMC321株和日本野猪源GUB-71株,其相似性为96.0%、95.1%和97.0%.该TM-a株轮状病毒不同基因表现出与人源轮状病毒和猪源轮状病毒的高度同源性,推测猪A群轮状病毒可能在人畜间传播,发生基因重组现象.

关 键 词:猪A群轮状病毒  分离  鉴定

Isolation and Identification of Group A Porcine Rotavirus
KU Xu-gang , ZHANG Kun , LIU Yu-xi , HE Qi-gai.Isolation and Identification of Group A Porcine Rotavirus[J].Acta Veterinaria et Zootechnica Sinica,2012,43(2):275-281.
Authors:KU Xu-gang  ZHANG Kun  LIU Yu-xi  HE Qi-gai
Institution:1,2 (1.State Key Laboratory of Agriculture Microbiology of Animal Virology,Wuhan 430070, China;2.College of Animal Science and Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China)
Abstract:Porcine rotavirus can induce serious diarrhea in piglet,but most researches are on human.We successfully isolated and identified group A porcine rotavirus(GAR) virus,which make a contribution to pathogenesis,molecular biology study on porcine rotavirus.GAR were detected by RT-PCR test from different feces samples of diarrhea,those positive samples were treated by trypsin,then was inoculated to MA-104 cell line to isolate strain TM-a.After routine RT-PCR test and electron microscope observation,we sequenced and analyzed VP6,VP7 and VP8 genes of strain TM-a.Through homology analysis,we found VP6,VP7 and VP8 are,respectively,identical to Beijing human strain LL3354 from China(96.0%),Indian human strain RMC321(95.1%),wild boar strain GUB-71 from Japan(97.0%).Strain TM-a have a high similarity to human and wild boar rotavirus,we infer that GAR may spread from human to animal,and there exist gene recombination.
Keywords:group A porcine rotavirus  isolation  identification
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