猪流行性腹泻病毒变异株分离鉴定及攻毒模型建立  

Isolation and identification of porcine epidemic diarrhea virus variant strain and establishment of challenge model

作　　者：焦安琪 郭效珍 陈智[2] 于江[2] 于克戬 逯晓寒 辛忠昊 韩先杰 吴家强 JIAO Anqi;GUO Xiaozhen;CHEN Zhi;YU Jiang;YU Kejian;LU Xiaohan;XIN Zhonghao;HAN Xianjie;WU Jiaqiang(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China;Key Laboratory of Animal and Poultry Disease Control and Breeding,Shandong Province,Institute of Animal Husbandry and Veterinary Medicine,Shandong Academy of Agricultural Sciences,Jinan 250000,China;Key Laboratory of Poultry Disease Diagnosis and Immunity in Shandong Province,Poultry Research Institute,Shandong Academy of Agricultural Sciences,Jinan 250000,China)

机构地区：[1]青岛农业大学动物医学院,青岛266109 [2]山东省农业科学院畜牧兽医研究所山东省畜禽疫病防治与繁育重点实验室,济南250000 [3]山东省农业科学院家禽研究所山东省家禽疫病诊断与免疫重点实验室,济南250000

出　　处：《生物学杂志》2023年第3期69-73,共5页Journal of Biology

基　　金：山东省现代农业产业技术体系项目(SDAIT-08-01);山东省农业重大应用技术创新项目(SD2019XM007);高层次人才工程项目;山东省农业科学院创新工程项目;国家自然科学基金项目(32070178);山东省重点研发计划项目(农业良种工程)。

摘　　要：猪流行性腹泻(PED)是由猪流行性腹泻病毒(PEDV)引起的一种急性高度接触性肠道传染病。研究成功分离到一株PEDV,命名为PEDV-20200946。对该毒株的S基因进行测序分析,遗传进化树结果显示,该分离毒株属于G2亚群。通过Reed-Muench法测定该毒株毒价为106.1 TCID 50/mL。通过动物回归试验进一步探究该分离毒株对仔猪的致病情况,将12头20日龄仔猪平均分为3组,分别为口服攻毒组、口服+肌注攻毒组和对照组。攻毒后观察临床症状,72 h后采集各组的粪便及发病猪的肠道进行RT-PCR检测和免疫组化分析。结果显示,口服+肌注组仔猪发病最快,口服攻毒组次之,对照未发病。剖检攻毒组仔猪可见其肠壁变薄、肠道膨胀,内含大量黄色液体。免疫组化试验显示,攻毒组猪小肠内可检测到大量PEDV阳性信号,而对照组未检测到。研究建立了PEDV-20200946毒株仔猪攻毒模型,为深入研究PEDV流行毒株的分子特性以及弱毒疫苗的开发奠定基础。Porcine epidemic diarrhea(PED)is an acute contact intestinal infectious disease in pigs caused by the porcine epidemic diarrhea virus(PEDV).In this study,a PEDV strain was isolated and named PEDV-20200946.S protein gene was sequenced and analyzed,and the phylogenetic tree showed that the isolated strain belonged to G2 genotype.The virus titer of this strain was 106.1 TCID 50/mL determined by Reed-Muench method.Subsequently,the pathogenicity of the isolated strains on piglets was further explored through animal challenge.A total of twelve 20-day-old piglets were divided into three groups on average,and they were named oral group,oral and intramuscular injection group,and control group,respectively.After the viral infection,clinical symptoms were observed,and the feces of each group and the intestines of the affected pigs were collected 72 h later for RT-PCR detection and immunohistochemical analysis.The results showed that piglets in the oral and intramuscular injection group showed the fastest diarrhea occurrence,followed by the oral group,and the control did not have any clinical symptoms.Autopsy examination showed that the intestinal walls of diseased piglets got thinned,their intestines were swollen,and contained a large amount of yellow liquid.Immunohistochemical tests showed that a large number of PEDV positive signals could be detected in the small intestine of piglets in the challenge groups,while the control group showed normal.The piglet challenge model of PEDV-20200946 strain was established,which laid a foundation for in-depth study of the molecular characteristics of PEDV epidemic strains and the development of attenuated vaccines.

关 键 词：猪流行性腹泻病毒 流行毒株 分离鉴定 攻毒模型 遗传进化树 

分 类 号：S828[农业科学—畜牧学] S852.65[农业科学—畜牧兽医]