2018年江西及福建省新现猪急性腹泻冠状病毒的分子流行病学调查  

Molecular Epidemiology of Newly Emerged Swine Acute Diarrhea Syndrome Coronavirus in Jiangxi and Fujian Provinces in 2018

作　　者：张誉瀚 袁为锋 张帆帆[1,2] 李凯 李指全[1,2] 宋德平 唐玉新 ZHANG Yuhan;YUAN Weifeng;ZHANG Fanfan;LI Kai;LI Zhiquan;SONG Deping;TANG Yuxin(Department of Preventive Veterinary Medicine,College of Animal Science and Technology,Jiangxi Agricultural University,Nanchang 330045,China;Key Laboratory for Animal Health of Jiangxi Province,Nanchang 330045,China)

机构地区：[1]江西农业大学动物科技学院,江西南昌330045 [2]江西省动物疫病诊断与防控重点实验室,南昌330045

出　　处：《养猪》2019年第4期113-117,共5页Swine Production

基　　金：国家重点研发计划项目(2017YFD0500600);江西省教育厅科技计划项目(GJJ160399)

摘　　要：猪急性腹泻综合征冠状病毒(Swine Acute Diarrhea Syndrome Coronavirus,SADS-CoV)是2017年在我国广东首次发现的一种可导致新生仔猪急性水样腹泻及死亡的新型冠状病毒。为调查2018年江西及福建猪群中该病毒的感染情况,试验建立了一种SADS-CoV套式RT-PCR检测方法,并调查2018年江西及福建腹泻猪只样本中SADS-CoV的流行情况。根据GenBank数据库中SADS-CoV GDS04毒株(登录号MF167434)的核衣壳蛋白(N)基因核酸序列,设计了两对特异性套式引物。以猪流行性腹泻病毒、猪传染性胃肠炎病毒、猪繁殖与呼吸综合征病毒、猪瘟病毒等病毒的基因组为模板检测引物的特异性。通过优化反应条件建立了检测SADS-CoV的套式PCR方法,用建立的套式PCR方法对2018年从江西和福建两省8个猪场采集的492份腹泻样品进行检测。应用设计的两对套式引物扩增出了650 bp和291 bp两个特异性片段。试验建立的套式PCR检测方法特异性好,只能扩增SADS-CoV,其最低检测限度为102拷贝/μL。利用所建立的方法对来自江西和福建省的492份腹泻猪临床样品进行检测,从福建猪群样品中检出了2份SADS-CoV感染,未从江西临床样品中检出SADS-CoV。试验将为SADS-CoV的临床检测提供一种套式PCR检测方法,结果表明该病毒在福建猪群而未在江西腹泻猪群中流行,将对SADS-CoV的临床诊断等具有应用价值。Swine acute diarrhea syndrome coronavirus(SADS-CoV)is a new coronavirus associated with acute watery diarrhea and death in newborn piglets,which was first found in Guangdong province of China in 2017.To investigate the presence and prevalence of this virus in pigs in Jiangxi and Fujian provinces in 2018,a nested RT-PCR assay would establish and used to investigate the SADS-CoV in clinical samples from pigs in the two provinces.Two pairs of specific primers were designed based on the nucleocapsid(N)gene sequence of the published SADS-CoV strain GDS04(GenBank accession number MF167434).The specificity of the primers was verified by using genomes of SADS-CoV,porcine epidemic diarrhea virus,transmissible gastroenteritis virus,porcine reproductive and respiratory syndrome virus,and classical swine fever virus as templates.A nested PCR assay for detecting SADS-CoV was established by optimizing the reaction conditions.A total of 492 clinical samples from pig farms in Jiangxi and Fujian provinces in 2018 were detected by using the established nested RT-PCR asssy.Two specific fragments with sizes of 650 base pair(bp)and 291 bp were amplified as expected.The nested PCR assay established in this experiment is specific and could only amplify SADS-CoV with a detection limit of 102 copies/μL.The established assay was used to detect 492 clinical samples of pigs from Jiangxi and Fujian provinces,and 2 samples were positive for SADS-CoV from Fujian swine population samples,and no sample was found positive for SADS-CoV in Jiangxi clinical samples.In this study,we have afford a nest RT-PCR for detection of SADS-CoV,and our results showed that the virus was existed in pigs in Fujian province and not prevalent in pigs in Jiangxi province,and it would have application value in the clinical diagnosis of SADS-CoV.

关 键 词：猪急性腹泻综合征冠状病毒 套式PCR 腹泻 分子流行病学 

分 类 号：S855.3[农业科学—临床兽医学]