猪瘟病毒RT-LAMP检测方法的建立  

Establishment of RT-LAMP system for rapid detection of classical swine fever virus

作　　者：李健[1,2] 熊炜[2] 方雪恩[3] 王巧全[2] 姜平[1] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]上海出入境检验检疫局,上海200135 [3]上海大学生命科学学院,上海200444

出　　处：《中国兽医科学》2010年第10期1033-1038,共6页Chinese Veterinary Science

基　　金：科技部科技支撑计划世博科技专项(2009BAK43B30)

摘　　要：利用逆转录环介导等温核酸扩增技术(RT-LAMP),建立了猪瘟病毒快速检测方法,同时评价了该方法的灵敏性和特异性。结果,根据猪瘟病毒5′端非编码区的一段保守序列设计的LAMP引物能够在65℃下1h内实现目标核酸区段的大量扩增,检测结果可直接用肉眼判断。结果表明,该检测体系具有极高的特异性,只能检测到目标病毒,与其他类似病毒如猪繁殖与呼吸综合征病毒、伪狂犬病病毒、传染性胃肠炎病毒、猪细小病毒和猪圆环病毒2型的核酸等无交叉反应,可检测到1×10-3稀释度的目标病毒核酸量,比普通RT-PCR的灵敏性高10倍。A rapid diagnostic method for classical swine fever（CSF）was established by using reverse transcription loop-mediated isothermal amplification（RT-LAMP）,meanwhile its specificity and sensitivity were assessed.The classical swine fever virus（CSFV）RNA could be amplified by incubation at 65℃ for only 1husing six special primers designed based on CSFV 5′non-coding sequence gene and the amplification products could be observed easily by naked-eye.The results showed that the developed system was specific for the detection of CSFV,and there was no cross-reaction with porcine reproductive and respirato- ry syndrome virus,pseudorabies virus,transmissible gastroenteritis virus,porcine parvovirus or porcine circovirus type 2（PCV-2）.The detection limit of the system was found to be 1×10-3 RNA sample,which was 10-fold higher than that of the traditional PCR.

关 键 词：猪瘟病毒 逆转录环介导等温核酸扩增 检测 

分 类 号：S855.3[农业科学—临床兽医学] S852.651[农业科学—兽医学]