鸡痘病毒PCR检测方法的建立  

Application of Polymerase Chain Reaction (PCR) for Detection of Fowlpox Virus Infection

作　　者：郭建顺[1] 金宁一[1] 夏志平[1] 金扩世[1] 徐敬龙[1] 贾雷立[1] 王凯[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2004年第6期534-536,共3页Chinese Journal of Veterinary Science

基　　金：国家"8 63"计划资助项目 ( 10 1-0 5 -0 3 -0 1) ;军队青年基金资助项目 ( 0 1Q12 7)

摘　　要：根据已发表的鸡痘病毒 (fowlpox virus,FPV) 4 b核心蛋白基因的核苷酸序列设计合成了 2对引物 ,通过对影响PCR扩增因素的优化 ,2对引物在同一反应条件下 ,以抽提 FPV DNA或直接用其毒液制备的模板均能分别扩增出预期的 5 4 9、136 1bp的片段。特异性检测表明 ,2对引物对 FPV10 2株、FPV2 82 E4 (北京 )、FPV2 82 E4 (吉林 )、v FV2 82重组鸡痘疫苗毒及 FPV临床分离株均能扩增出相应特异性片段 ,而用鸡马立克氏病病毒、鸡传染性喉气管炎病毒、羊痘病毒、鸡胚成纤维细胞 (CEF)制备的模板分别进行 PCR扩增却成阴性。敏感性检测表明 ,2对引物 (p1 ,2 、g1 ,2 )分别能检测到 10 - 2 、10 - 3fmol的 FPV DNA和 10 0 .5、10 - 0 .5TCID50 的病毒。以上结果表明 ,本试验所建立的 FPV PCR检测法具有较高的特异性和敏感性 ,模板制作简单 ,完全可用于In order to establish a more rapid ,sensitive and special method for detection of fowlpox virus (FPV), two pairs of primers(p_(1,2), g_(1,2)) were designed and synthesized on the basis of published DNA sequence of the 4b core protein gene of the FPV genome in this study. The PCR method was developed for detection of FPV infection.The results revealed the expected 549 bp or 1 361 bp fragment could be amplified respectively by two sets of primers from extracted DNA or virus suspensions from recombinant FPV, FPV vaccine strains or FPV field isolates. There was no amplification in control samples e.g., Marek′s disease virus (MDV), avian infectious laryngotracheitis virus(AILTV), goatpox virus, DNA from chicken embryo fibroblast cell (CEF) using any of the primers. Two pairs of primers of this method could detect at least 10^(-2) fmol(p_(1,2)), 10^(-3) fmol(g_(1,2)) FPV DNA and the virus of 10^(0.5)(p_(1,2)), 10^(-0.5) (g_(1,2)) TCID_(50), respectively. The results showed the PCR technique had the specificity and high sensitivity, and would be applied for diagnosis of fowlpox virus infection.

关 键 词：鸡痘病毒 PCR检测方法 引物设计 模板制备 

分 类 号：S858.31[农业科学—临床兽医学]