猪博卡病毒PCR检测方法的建立及其初步流行病学调查  

Establishment of the PCR assay for porcine Bocavirus detection and primary epidemiology survey of the virus infection

作　　者：胡军勇[1] 张倩[2,3] 王丹丹[2] 涂志勤[2] 胡睿铭[2] 汤细彪[2] 吴斌[2,3] 

机构地区：[1]华中农业大学动物科技学院,湖北武汉430070 [2]华中农业大学动物传染病诊断中心,湖北武汉430070 [3]农业微生物学国家重点实验室,湖北武汉430070

出　　处：《中国预防兽医学报》2012年第8期632-636,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：湖北省研究与开发计划项目(201OBBB08)

摘　　要：猪博卡病毒(PBoV)是一种新出现的病毒,可以导致仔猪急性腹泻。为建立该病毒的检测方法,本实验根据PBoV的NS1基因序列设计一对引物,建立了PCR检测方法。特异性试验结果表明,该方法能够从PBoV阳性样品中特异性的扩增出482 bp的片段,而对猪伪狂犬病毒、猪细小病毒、猪细小病毒、猪圆环病毒、沙门氏菌、大肠杆菌、猪链球菌、副猪嗜血杆菌的核酸样品均无非特异性扩增反应。敏感性试验显示,该PCR方法对PBoV的最低检测量为213.6拷贝。此外,应用该方法对湖北、湖南、河南省的各大猪场进行了流行病学调查,在79个规模化猪场采集的248份病料样品中有172份样品为PBoV阳性。其中,对4个发生腹泻疫情的规模化猪场进行了分群抽样调查,结果显示PBoV在腹泻猪群中的阳性率达到73.95%,并显著高于非腹泻猪群(47.83%)。本研究调查结果显示PBoV在国内猪群中流行广泛而且感染率高。Porcine Bocavirus （PBoV） is newly emerged in China which is supposed to be associated with the cause of diarrhea in piglets. To establish the assay for PBoV detection, a PCR protocol was developed with a pair of primers designed according to NS1 gene available in GenBank. Under the optimized conditions of the PCR reaction, the specific fragment of 482 bp was amplified from the PBoV NS1 gene containing recombinant plasmid with a limit detection equivalent to 213.6 copies, but no amplifications were found from genome DNA of pseudorabies virus, porcine parvovirus, porcine circovirus type 2, Salmonella, E. coil, S. suis, H. parasuis. Furthermore, A total of 248 clinical samples were collected from pig herds with unexplained diarrhea in Hubei, Hunan and Henan provinces, and 172 samples of which were PBoV positive （69.35%） detected by the established PCR assay. In additionally, faeces samples were collected from different age groups of pigs in 4 pig farms with diarrhea and the detection results showed that PBoV positive rate among diarrhea pig （73.95%） was significantly higher than that of the pig groups without diarrhea （47.83%）, indicating that PBoV infections might be associated with the causes of diarrhea prevalence in piglets.

关 键 词：猪博卡病毒 PCR检测方法 腹泻 流行病学调查 

分 类 号：S852.65[农业科学—基础兽医学]