猪博卡病毒1型/2型PCR检测方法的建立  

Establishment of PCR for detection of porcine Bocavirus type 1/2

作　　者：付朋飞[1,2] 李梦奇[3] 乔涵[1] 杨兴武[1] 张宇[1] 徐朋丽 韩昊莹 张鸿鑫 陈红英[1] 

机构地区：[1]河南农业大学郑州市猪重大疫病防控重点实验室,河南郑州450002 [2]北京大北农动物医学研究中心,北京100195 [3]华南农业大学兽医学院,广东广州510642

出　　处：《中国预防兽医学报》2016年第7期572-575,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：河南省重大科技专项(111100110300)

摘　　要：猪博卡病毒(PBo V)是2009年新发现的一种DNA病毒,常于腹泻仔猪体内检测到该病毒。为建立该病毒的检测方法,本研究根据Gen Bank中登录的PBo V 1/2型的高度同源保守序列设计一对特异性引物,经PCR扩增条件优化,建立了PBo V 1/2型的PCR检测方法。特异性试验结果显示,该PCR方法仅从PBo V 1或2型的阳性病料样品中特异性的扩增出751 bp的目的片段,而对猪伪狂犬病毒、圆环病毒、大肠杆菌、沙门氏菌的核酸样品的扩增结果均为阴性。敏感性试验显示,该PCR方法对PBo V 1/2重组质粒标准品的最低检测量为34.8拷贝/μL。对来自河南中牟、开封、兰考、濮阳、洛阳等地猪场发生腹泻疫情的70份猪病料样品,采用该方法进行检测,其中10份样品呈PBo V 1/2型阳性,阳性率为14.29%。该PCR检测方法的建立为PBo V1/2型的诊断和监测提供技术支持。Porcine Bocavirus （PBoV） is a newly emerging DNA virus in 2009 which was supposed to be associated with the diarrhea in piglets. To establish the assay for PBoV detection, a pair of primers were designed and synthesized according to the highly homologous regions of published PBoV1/2 sequences available in GenBank. Thus, a PCR protocol for PBoV1/2 detection was developed under the optimized conditions for the amplification. The specific fragment of 751 bp was amplified only from the template DNA of PBoV1/2, but no amplifications were found from DNAs for other pathogens such as pseudorabies virus, porcine circovirus type 2, Salmonella or E.coli. The PCR method was capable of detecting the template DNA of 34.8 copies/μL. A total of 70 clinical samples collected from several areas from Zhongmou, Kaifeng and Puyang were detected by the established PCR assay, and 10 samples were positive for PBoV1/2 （14.29%）. These results indicated that the PCR method could be application in PBoV1/2 detection in the virus infected pigs.

关 键 词：猪博卡病毒 PCR检测方法 腹泻 

分 类 号：S852.65[农业科学—基础兽医学]