区分马立克氏病病毒疫苗株CVI988与其他毒株的PCR方法的建立及应用  

Establishment and employment of PCR technique for distinguish Marek′s disease virus vaccine strain CVI988 from other MDV strains

作　　者：丁家波[1] 姜世金[1] 朱鸿飞[2] 崔治中[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]中国农业科学院生物制品工程技术中心,北京100081

出　　处：《中国兽医学报》2007年第1期39-42,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金重点资助项目(30300450);国家自然科学基金资助项目(30070544)

摘　　要：利用马立克氏病病毒(MDV)疫苗毒CVI988株pp38基因上游启动子区域连续5个碱基的缺失(5′-AGCCG-3′),设计2对特异性引物,建立了能区分MDVCVI988株和MDV其他毒株的PCR诊断方法。该方法对8株MDV毒株(弱毒疫苗株CVI988和814,强毒参考株GA,超强毒参考株RB1B、Md5和Md11,特超强毒参考株648A及1个中国野毒株J-E)的PCR鉴定结果均与预期吻合。以16只疑为MDV感染鸡的脏器组织核酸提取物为模板,用所建立的PCR方法,能从其中7只鸡样品中扩增出特异性条带,其检测结果与细胞分离培养和单克隆抗体的检测结果一致。试验证实,本研究所建立的PCR诊断方法具有良好的特异性和敏感性。The promoter of pp38 gene in Marek＇s disease virus（MDV） vaccine strain CVI988 had a consecutive 5 bp （5＇-AGCCG-3＇）deletion. In this study, two pairs of primers were designed and synthesized to distinguish CVI988 strain from other MDV strains by polymerase chain reaction （PCR）, 8 MDV strains （vaccine strains CVI988 and 814, virulent reference strains GA, very virulent reference strains RB1B, Md5 and Md11,very virulent plus reference strains 848A,and a Chinese field strain J-E） were distinguished by this technique and the detected result was consisted with the putative. Chicken tissue nucleic acid extracts from MDV-suspected chickens were used as the templats for PCR, and 7 samples out of 16 were MDV positive in the detection. At the same time, lymphocytes were isolated from the sixteen chickens for cell culture in chicken embryo fibroblast （CEF）, respectively. In the second passage, only 7 samples of the PCR positive produced MDV specific plaque, the other 9 samples could not even when the cells were passed to the fifth passage. The cultivated cells were further detected by MDV specific monoclonal antibodies and confirmed the result of PCR. The above results indicated that the established PCR in this study was sensitive enough to distinguish MDV CVI988 strain from other MDV strains.

关 键 词：马立克氏病病毒 疫苗毒CVI988 PCR 

分 类 号：S852.65[农业科学—基础兽医学]