四川部分地区新生仔猪病毒性腹泻的病原学诊断  

Etiologic Diagnosis of Viral Diarrhea in newborn piglets from partial Sichuan Area

作　　者：高继业[1] 黄伟[1] 王振兴[1] 庞璞 温亚兰 谢立飞[1] 

机构地区：[1]西南大学荣昌校区,四川荣昌402460

出　　处：《中国兽医杂志》2014年第1期9-13,共5页Chinese Journal of Veterinary Medicine

基　　金：西南大学青年基金项目(2010RCQ002)

摘　　要：根据参考文献合成5对针对猪传染性腹泻病毒(PEDV)M基因、猪传染性胃肠炎病毒(TGEV)S基因、猪轮状病毒(PRV)VP4基因、博卡病毒(PBoV)VP1/VP2基因、猪库布病毒(PKoV)3D基因,可分别特异扩增出大小为467 bp、1 062 bp、750bp、496bp和323 bp的DNA片段的特异性引物,对四川部分地区2011年10月至2012年5月新生仔猪腹泻病例样本进行RT-PCR、PCR检测和透射电镜观察。检测结果显示,SC1、SC2猪场的病例样本均扩增出PKoV 3D基因特异性DNA片段,均未扩增出TGEV、PRV、PBoV的特异性DNA片段,仅SC1猪场有1份样本扩增出PEDV M基因的特异性DNA片段。测序结果表明,扩增出的PKoV 3D基因特异性DNA片段大小为323 bp,与GenBank登录序列同源性均在95%以上;扩增出的PEDV M基因特异性DNA片段大小为467 bp,与GenBank登录序列同源性均在98%以上。透射电镜观察结果显示,两个猪场样本均可见有直径大小为25、30 nm的球形病毒颗粒。结果显示,此次四川部分地区猪场发生的病毒性腹泻病例均存在PKoV感染,说明PKoV与此次疫情有紧密联系。According to the references, five pairs of primers for the amplication of M gene of Porcine epidemic diarrhea virus （PEDV） , the S gene of Transmissible gastroenteritis virus （TGEV）, the VP1 / VP2 gene of Porcine rotavirus （PRV） ,the VP4 gene of Porcine Bocavirus （PBo V） ,the 3D gene of Porcine Kobuvirus （PKo V） were synthesized, and the products had molecular sizes of 467 bp , 1062 bp , 750 bp , 496 bp and 323 bp respectively. The primers were used to detect PEDV ,TGEV ,PRY ,PBoV ,PKoV in cases of newborn piglets diarrhea from partial Sichuan area from October 2011 to May 2012 by RT-PCR,PCR and transmission electron microscope. The RT-PCR / PCR results showed that PKoV were tested, and TGEV, PRY ,PBoV were not detected in all samples except PEDV detected tested in one sample. The results of sequencing and analysis showed that the DNA fragment of PKoV 3D gene, PEDV M gene had size of 323 bp , 467 bp and shared more than 95 % ,98 % homology respectively with the sequence logined in GenBank. Spherical virus particles with the size of 25-30 nm were found in all samples by transmission electron microscope. The results clearly indicated that PKo V were distributed in all case, which suggests that positive relationship exists between this diarrhea disease outbreak and porcine kobuvirus .

关 键 词：仔猪腹泻 猪传染性胃肠炎病毒 猪流行性腹泻病毒 猪轮状病毒 猪博卡病毒 猪库布病毒 

分 类 号：S858.28[农业科学—临床兽医学]