猪伪狂犬病病毒LAMP可视化检测方法的建立  

Establishment of a Visualized LAMP Method for Detection of Pseudorabies Virus

作　　者：许宗丽[1] 黄溢泓[1] 李志源[1] 刘针伶[1] 周师师 覃艳然 邱洁 盘美妮[1] Xu Zongli;Huang Yihong;Li Zhiyuan;Liu Zhenling;Zhou Shishi;Qin Yanran;Qiu Jie;Pan Meini(Liuzhou Animal Disease Prevention and Control Center,Liuzhou,Guangxi 545000)

机构地区：[1]柳州市动物疫病预防控制中心,广西柳州545000

出　　处：《中国动物检疫》2017年第10期79-82,共4页China Animal Health Inspection

基　　金：广西柳州科技项目(2015E040501)

摘　　要：根据Gen Bank中猪伪狂犬病病毒(PRV)g B基因的保守序列,设计一套特异性引物,在对反应条件进行优化后,建立了PRV环介导等温扩增(LAMP)可视化检测方法。结果显示:该方法能在常规65℃水浴锅中30 min内实现对目的片段的特异性扩增,并且可通过肉眼观察颜色直接判定检测结果 ;该方法具有很高的特异性,与猪瘟病毒、猪圆环病毒2型等均无交叉反应;该方法的敏感性可达1 fg;将该方法应用于临床样品检测,结果与普通PCR方法一致。研究表明,本研究建立的LAMP方法简便、快速、灵敏、特异,适合基层PRV感染的快速检测。According to the sequences of PRV gB gene in GenBank,a set of primers was designed.After the optimizationof the reaction conditions,a visualized LAMP for PRV was established.The results showed that the optimizeddetection assay could be conducted by incubating in the conventional water bath at65℃for30min,and the result couldbe directly determined by visual observation.The assay was specific to PRV with a detection limit of1fg,and no crossreactivitywith other pathogens of swine,including CSFV and PCV2.The results of detection of clinical samples byour LAMP approach were consistent with that of the ordinary PCR assay.These results suggested that the LAMP assaydescribed in this study was a simple,rapid,specific method for detection of PRV in the field.

关 键 词：猪伪狂病病毒 环介导等温扩增(LAMP) 检测 

分 类 号：S851.3[农业科学—预防兽医学]