牛白血病病毒LUX^TM实时荧光聚合酶链式反应检测方法的建立  

Establishment of LUX^(TM) real-time fluorescent PCR/RT-PCR for rapid detection of bovine leukemia virus

作　　者：刘志玲[1] 陈茹[2,1] 马静云[1] 吴晓薇[2] 曾碧健[2] 林志雄[2] 

机构地区：[1]华南农业大学动物科学院,广东广州510642 [2]广东出入境检验检疫局,广东广州510623

出　　处：《中国预防兽医学报》2009年第5期383-387,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局科研项目(2006IK-019)

摘　　要：采用LUXTM新型荧光PCR技术原理,设计并合成1对单标记LUXTM荧光引物,建立了LUXTM荧光PCR和RT-PCR方法,可快速检测牛白血病病毒(BLV)前病毒DNA和病毒RNA。结果显示所建立的LUXTM荧光PCR/RT-PCR体系可特异检测牛白血病病毒核酸,而对蓝舌病、牛病毒性腹泻-粘膜病、水泡性口炎、口蹄疫、牛流行热、牛传染性鼻气管炎等病毒核酸以及牛结核分支杆菌、大肠杆菌、健康动物组织核酸扩增均呈阴性反应。敏感性实验表明LUXTM荧光RT-PCR对RNA前病毒的检测敏感性达0.49 ng/μL,荧光PCR对pTblv-gp51重组质粒的检测敏感性可达0.138拷贝,比OIE规程的巢式PCR方法高104倍以上。采用该方法从血清学阳性临床牛血清中检出BLV阳性样品。A LUXTM fluorescent RT-PCR/PCR assay for detecting Enzootic bovine leukemia virus （BLV） was established using specific primers based on gp51 gene sequence. The specificity of the assay was confirmed against BLV and related bovine virus （BTV, BVDV, VSV, FMDV, BEFV）, M. tuberculosis, Escherichia coli and genomic DNA from healthy cattle. The RT-PCR assay had a detection limit of 0.49 ng/μL RNA and could detect as few as 0.138 copies of pTblv-gp51 recombinant plasmid, which was 104-fold more sensitive than Nested PCR method.

关 键 词：牛白血病病毒 荧光PCR LUXTM引物 

分 类 号：S851.34[农业科学—预防兽医学]