3种苹果潜隐性病毒一步多重RT-PCR检测体系的优化

为建立更为快速准确的苹果潜隐性病毒检测方法,以携带苹果茎沟病毒(ASGV)、苹果褪绿叶斑病毒(ACLSV)和苹果茎痘病毒(ASPV)的田间苹果成龄树为试材,设计合成了扩增目的片段为273 bp(ASGV)、358bp(ACLSV)和432 bp(ASPV)的3对特异性引物,并对影响RT-PCR体系的主要参数进行试验优化.结果表明:建立的一步多重RT-PCR体系可以实现对3种主要潜隐性病毒的特异性检测；通过对田间多个样品的检测验证,表明该方法的准确性和灵敏度都很高,并缩短了单个样品的检测时间,极大地提高了工作效率.

Quick detection of three apple latent viruses by one-step multiplex RT-PCR

In order to establish a rapid and reliable method to detect three apple latent viruses,mature apple trees infected by Apple stem grooving virus(ASGV),Apple chlorotic leaf spot virus(ACLSV) and Apple stem pitting virus(ASPV) were taken as materials,and three sets of specific primers with fragments of 273 bp(ASGV),358 bp(ACLSV) and 432 bp(ASPV) were amplificated.Some main parameters affecting the RT-PCR system were tested.The results showed that one-step multiplex RT-PCR could specially detect the three apple latent viruses.The sensitivity and applicability were verified by testing many samples from field.The detection of the three apple viruses from a single sample can be finished effectively and can improve the efficiency greatly.