利用RT-PCR快速检测马铃薯X病毒(PVX)

Rapid Detection of Potato Virus X by RT-PCR

采用TR IZOL试剂盒和异硫氰酸胍法2种方法提取了带PVX马铃薯试管苗的总RNA。根据设计好的1对特异性引物,运用反转录PCR技术对提取的总RNA进行了体外扩增。结果表明,所用2种方法都能有效地提取马铃薯RNA,并适合于反转录合成病毒cDNA,得到了与预期大小相一致的720 bp片段,而对照未得到任何产物。但利用TR IZOL试剂盒进行RNA提取价格较贵,而异硫氰酸胍法所用试剂可自己配制,方法灵活,价格较TR IZOL试剂盒便宜。从而建立了经济简便的PVX RT-PCR检测体系,为PVX的防治、脱毒种薯和核心种苗的检测提供有效手段。

The potato virus X genomic RNA was obtained from PVX-infected potato in vitro by two methods of TRIZOL kit and Guanidine Iso- thiocyanate Method （ GIM ）. A pair of specific primers were designed and synthesized. The results showed that all the two methods could extracted potato RNA effectively. By the reverse transcription and polymerase chain reaction （ RT-PCR）, a 720bp DNA fragment was amplifyied successfully from PVX-RNA and no fragment was obtained from the control. But the prices of using TRIZOL kit to extrated RNA is more expensive, while the Guanidine Isothiocyanate Method can be prepared reagent by self flexibility, and its price cheaper than the TR- IZOL kit. So, an economic and convenient RT-PCR detection system of PVX was set up, and provided an effective means of detection for the prevention and treatment PVX, the production of potato virus-flee seeds and core plants.