内源和外源性禽白血病病毒鉴别PCR检测方法的建立  

Establishment of a PCR method for differentiating endogenous and exogenous avian leukosis virus

作　　者：周刚[1] 牛成明 司昌德[1] 韩凌霞[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2010年第7期546-549,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：中央级公益性科研院所基本科研业务费项目(2009-08)

摘　　要：为建立简单快捷的内、外源性禽白血病病毒(ALV)的鉴别检测方法,本研究根据外源性ALVA亚群标准株RAV-1的p27、env基因以及内源性ALVE亚群标准株ev1的env基因的保守序列,设计3对特异性引物,可分别对ALV、外源性ALV(A、B亚群)和内源性ALV(E、J亚群)进行扩增,扩增产物分别为793bp、387bp和234bp,通过对各反应条件的优化建立了同时检测并鉴别内、外源性ALV的多重PCR方法。该方法特异性良好、灵敏度可达到2×103copies,利用该方法和ELISA对5份现地病鸡组织样品和10枚疑似感染内源性ALV的鸡胚进行检测,结果表明:4份病鸡样品均扩增出3条特异性片段;而9枚鸡胚仅扩增出793bp和234bp的特异性片段,2种检测方法的符合率为100%。该方法为内、外源性ALV的临床鉴别诊断奠定了基础。A multiplex PCR for differentiating exogenous and endogenous avian leukosis virus（ALV） was developed using primers derived from p27 and env gene of ALV subgroup A and env gene of ALV subgroup B.This method was shown to specifically amplify 793 bp,387 bp and 234 bp PCR products from exogenous and endogenous ALVs of all subtypes.The multiplex PCR was evaluated by test on 5 suspected clinical samples and 10 chicken embryos.All three PCR fragments were amplified from 4 clinical samples,indicating these were infected with both exogenous and endogenous ALV.However,only 793 bp and 234 bp PCR products were detected in 9 chicken embryos,suggesting that these embryos were infected only by endogenous ALV.The estamblished multiplex PCR assay would provide a rapid and sensitive detection method for ALV diagnosis.

关 键 词：禽白血病 外源性 内源性 复合PCR 

分 类 号：S852.659[农业科学—基础兽医学]