牛流行热病毒一步法RT-PCR检测方法的建立  

Establishment of One Step RT-PCR for Detecting Bovine Epizootic Fever Virus

作　　者：廖承球[1] 周庆丰[1] 陈俊伟[1] 潘永飞[1] 卢围[1] 王东东[1] 蔡新斌[1] 宋延华[1] 

机构地区：[1]广东温氏食品集团股份有限公司,广东新兴527400

出　　处：《动物医学进展》2014年第2期55-57,共3页Progress In Veterinary Medicine

摘　　要：根据NCBI上公布的牛流行热病毒G基因序列设计、合成了一对检测牛流行热病毒G基因的RT-PCR引物,以牛流行热疫苗为模板,建立了牛流行热的RT-PCR检测方法,经扩增得到大小799bp的片段。该方法具有良好的特异性、敏感性,能扩增出含量为9.576pg的BEFV RNA。从17份疑似牛流行热病毒感染的牛全血中检测出阳性病例5份,经测序后证明为牛流行热病毒G基因片段。结果表明,建立的一步法RT-PCR方法简便易行,可用于牛流行热的分离鉴定、临床病料的检测和分子流行病学调查。Based on G gene sequences of bovine epizootic fever virus published in NCBI, a pair of primers were designed and synthesized for detecting G gene. The 799 bp specific fragment was obtained from the BEFV vaccine strain RNA by one step RT-PCR. The results indicated that the one step RT-PCR reaction after optimization showed good sensitivity and specificity, the sensitivity of the method reached to 9. 576 pg. 5 samples of 17 samples suspected infection with BEFV were positive detecetd by one step RT-PCR. Results showed that the RT-PCR is specific and sensitive,and can be used as a method to identify BEFV i- solates, detect samples of clinical cases and investigate molecular epidemiology of BEFV.

关 键 词：牛流行热病毒 一步法RT-PCR 建立 

分 类 号：S852.659.5[农业科学—基础兽医学] Q789[农业科学—兽医学]