双引物探针RT-Realtime PCR检测马铃薯A病毒  

Detection of Potato virus A with double primers and probes RT-Realtime-PCR technique

作　　者：张京宣[1] 梁炜[1] 耿金培 陈洪俊[3] 杨益娥 粟智平 

机构地区：[1]山东出入境检验检疫局,山东青岛266000 [2]烟台出入境检验检疫局 [3]中国检验检疫科学研究院

出　　处：《植物检疫》2012年第1期26-28,共3页Plant Quarantine

基　　金：科技部"十一五"科技支撑计划(2006BAD08A13);国家质检总局科研项目(2006IK209)

摘　　要：马铃薯A病毒(Potato virus A,PVA)是我国重要的检疫性有害生物。本研究根据PVA中CP基因(coat protein gene)的保守序列,设计了两套PCR引物和TaqMan探针,建立了双引物探针RT-RealtimePCR检测PVA的方法。该方法采用实时荧光PCR技术,有效地提高了检测的灵敏度;同时两套引物探针相互验证,有效提高了结果的准确性。实验结果表明,本方法准确、灵敏、简便、快速,检出低限可达0.5fg/μL植物总RNA。Potato virus A（PVA） is one of the serious quarantine plant pests.Two suits of primers and Taqman probes were designed according to the coat protein gene of PVA in this study,and we set up a new method of RT-Realtime PCR for detection of PVA.The sensitivity was improved effectively by using the realtime PCR technique.At the same time two suits of PCR systems can test each other,which improves effectively the accuracy of the detection results.The method provides a accurate and sensitive detection of PVA,the limit content of the detection is 0.5fg/μL total plant RNA.

关 键 词：马铃薯A病毒 CP基因 双引物探针RT-Realtime PCR 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治] S41-30[农业科学—植物保护]