半巢式-RT-Realtime PCR检测李痘病毒  

Detection of Plum pox virus by semi-nested RT-Realtime PCR

作　　者：粟智平 张静 黄迎波[2] 李桂芬[3] 耿金培 朱水芳[3] 

机构地区：[1]烟台出入境检验检疫局,山东烟台264000 [2]湖南出入境检验检疫局 [3]中国检验检疫科学研究院

出　　处：《植物检疫》2012年第3期26-29,共4页Plant Quarantine

基　　金：国家质检总局科技计划项目(2010IK264);"十一五"科技支撑计划项目(2006BAD08A13);国家重点基础研究发展计划(2011CB932800)

摘　　要：李痘病毒(Plum pox virus,PPV)是我国重要的植物检疫性有害生物。本研究根据PPV中CP基因(coat protein gene)的保守序列,设计了3条PCR引物和1条TaqMan探针,建立了半巢式-RT-RealtimePCR检测PPV的方法。该方法有机地结合了巢式PCR和实时荧光PCR技术;3条引物形成的2套PCR体系相互验证,有效提高了结果的准确性;荧光探针有效提高了检测的灵敏度。实验结果表明,本方法准确、灵敏、简便、快速,检出低限可达37fg/μL植物总RNA。Plum pox virus （PPV） is one of the serious quarantine plant pests. Three primers and one TaqMan probe were designed according to the coat protein gene of PPV in this study, and we set up a new method of semi - nested RT - Realtime PCR for detection of PPV. The Nest - PCR and Reahime - PCR techniques were con- bined maneuverably in this method. Two suit of PCR systems made up with the three primers and one TaqMan probe can test each other ,which improves effectively the accuracy of the detection results. We improve effectively the sensitivity by using a fluorescence TaqMan probe. The method provides a accuracy and sensitive and easy and fast detection of PPV, and the limit content of the detection is 37fg/μL total plant RNA.

关 键 词：李痘病毒 外壳蛋白基因 半巢式-RT-Realtime PCR 

分 类 号：S41-30[农业科学—植物保护]