广西猪A群轮状病毒G9P[7]型的分离与初步鉴定  

Isolation and preliminary identification of group A porcine rotavirus type G9 P[7]from Guangxi

作　　者：米雪 陈荣琳 张胜斌 刘雪婷 秦秋英 王奕斐 边金妮 程振孔 罗允燕 罗期方 张宁 陈樱[1] 韦祖樟[1] 黄伟坚[1] 欧阳康[1] MI Xue;CHEN Ronglin;ZHANG Shengbin;LIU Xueting;QIN Qiuying;WANG Yifei;BIAN Jin-ni;CHENG Zhenkong;LUO Yunyan;LUO Qifang;ZHANG Ning;CHEN Ying;WEI Zuzhang;HUANG Weijian;OUYANG Kang(College of Animal Science and Technology,Guangxi University,Namiirig 530005,China;Yongxin Animal Husbandry Group Xijiang Co.LTD of Guangxi Agriculture Reclamation,Guigang,Guangxi 537104,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530005 [2]广西农垦永新畜牧集团西江有限公司,广西贵港537104

出　　处：《中国兽医学报》2021年第9期1722-1728,共7页Chinese Journal of Veterinary Science

基　　金：广西自然科学基金资助项目(2017GXNSFAA198138);广西大学学术骨干科研基金资助项目(XGZ170239);贵港市科学研究与技术开发计划资助项目(贵科攻1829012);“大学生创新创业训练计划”资助项目(201910593084)。

摘　　要：猪轮状病毒(porcine rotavirus,PoRV)感染仔猪可引起腹泻、脱水甚至死亡,是引起仔猪腹泻的常见病原,给养猪业造成巨大的经济损失。本研究为确定导致广西贵港某猪场仔猪腹泻的病原,采集腹泻仔猪的肠内容物,采用PCR方法检测猪流行性腹泻病毒(porcine epidemic virus,PEDV)、猪传染性胃肠炎病毒(transmissible gastroenteritis virus,TGEV)以及PoRV病原,结果显示,仅PoRV为阳性,将其接种至Marc-145细胞,盲传4代后,可产生细胞皱缩、脱落等明显细胞病变。稳定盲传20代,每隔5代进行PoRV的检测,选取第10代的病毒上清液进行VP4、VP6和VP7基因测序及遗传进化分析,确定该分离株为G9 P[7]型。电镜观察,可见到大小约为70 nm的轮状病毒粒子,符合轮状病毒特征,进一步鉴定分离株为PoRV。结果表明,PoRV的分离鉴定为其分子生物学特性研究以及该病的有效防控奠定了基础,也为疫苗的研发提供参考。Porcine rotavirus(PoRV)is a common pathogen of piglet diarrhea,PoRV infection in piglets can cause diarrhea,dehydration and even death,which causes great economic loss to the pig industry.In this study,intestinal contents of piglets with diarrhea were collected to determine the pathogens of diarrhea in a pig farm in Guigang,Guangxi and the pathogens of porcine epidemic virus(PEDV),transmissible gastroenteritis virus(TGEV)and PoRV were detected by RT-PCR.The results showed that only PoRV was positive.After inoculation to Marc-145 cells and blind transmission for four generations,cytopathic effects such as cell shrinkage and shedding were observed.Stable blind transmission was performed for 20 generations,and PoRV was detected every 5 generations.The 10 th generation isolates were selected for VP4,VP6 and VP7 gene sequencing and genetic evolution analysis,and the isolated strain was identified as G9 P[7].The wheel-like virions with a size of about 70 nm were observed by transmission electron microscopy,which was consistent with the characteristics of rotavirus,and the isolated strain was further identified as PoRV.The isolation and identification of PoRV lays a foundation for the study of molecular biological characteristics of PoRV and the effective prevention and control of PoRV,and provides reference for the research and development of vaccine in the future.

关 键 词：猪A群轮状病毒 腹泻 病毒分离 透射电镜观察 

分 类 号：S852.65[农业科学—基础兽医学]