采用PCR-RFLP法鉴别IBDV强毒BC6/85株和弱毒B87株  

Identification of IBDV virulence strains BC6/85 and vaccine strains B87 by PCR-RFLP

作　　者：陈仕怡 陈媛[1] 赖隆永 李春燕[1] 刘梦茜[1] 袁晓琴[1] 郑庆礼 许丽惠[1] 王全溪[1] CHEN Shiyi;CHEN Yuan;LAI Longyong;LI Chunyan;LIU Mengxi;YUAN Xiaoqin;ZHENG Qingli;XU Lihui;WANG Quanxi(College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学动物科学学院,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2019年第3期359-364,共6页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家星火计划重点项目(2015GA720001)

摘　　要：选取传染性法氏囊病病毒(IBDV)强毒力代表株BC6/85和弱毒力代表株B87为研究对象,根据VP2基因保守区设计一对通用引物和两个限制性内切酶位点,分别扩增两个毒株的VP2基因,然后进行BamHⅠ和PstⅠ酶切,采用限制性片段长度多态性聚合酶链反应(PCR-RFLP)分析酶切产物,并进行特异性、敏感性和重复性检测.结果表明:所设计的引物均可扩增两个毒株的VP2基因,大小约856 bp;经BamHⅠ和PstⅠ酶切后,BC6/85株的PCR产物被BamHⅠ和PstⅠ切出166、242和449 bp大小的3个片段,而B87株仅被PstⅠ切出242和615 bp大小的2个片段,且重复性好,特异性强.可见,采用PCR-RFLP分析IBDV VP2基因的方法操作简单,是一种能够快速鉴别IBDV强、弱毒株的可靠方法.In order to identify the virulent and attenuated strain of infectious bursal disease virus(IBDV),a pair of universal prim ers were designed to amplify the VP2 gene of IBDV strain BC6/85 and IBDV strain B87.Then the PCR products were digested by DNA restriction enzymes BamHⅠand PstⅠ.And AGE agarose gel electrophoresis was used to analyze the profile of enzyme diges ted products.Results showed that IBDV VP2 gene can be amplified by the general primer,with a target fragment being 856 bp.After restriction enzyme digestion,the PCR products of BC6/85 strain was cut into fragments of 166,242 and 449 bp,while the PCR products of B87 strain was only cut into fragments of 242 and 615 bp.PCR RFLP were proved a rapid method for identifying IBDV virulent and attenuated strain.

关 键 词：传染性法氏囊病病毒(IBDV) 限制性片段长度多态性聚合酶链反应(PCR RFLP) 限制性内切酶 鉴别诊断 

分 类 号：S852.65[农业科学—基础兽医学]