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 an important quarantine pest in our country. In this study, two sets of PCR primers and TaqMan probes were designed according to the conserved sequence of the coat protein gene in PVA. A double-primer RT-Realtime PCR method for the determination of PVA was established. The method uses real-time fluorescence PCR technique, which effectively improves the sensitivity of the detection; meanwhile, the two sets of primer probes mutually verify and effectively improve the accuracy of the result. The experimental results show that the method is accurate, sensitive, simple and rapid, and the detection limit is up to 0.5fg / μL of plant total RNA.