应用RT-PCR技术检测马铃薯A病毒

Detection of Potato Virus A by RT-PCR

参考GenBank中马铃薯A病毒(potato virus A,PVA)的保守序列,利用Primer6.0引物设计软件设计并合成了一对特异性引物PVAF、PVAR,以此引物利用RT-PCR方法对PVA保守序列基因进行了特异性扩增。结果表明:引物PVAF、PVAR能从已知的感染PVA病毒的植株中扩增出834bp的cDNA特异性片段;该RT-PCR的检测灵敏度为1pg的病毒核酸,特异性强,重复性好,可用于PVA病毒的快速检测。

A pair of primers PVAF and PVAR were designed according to the segmental nucleotide sequence of conservative gene of potato virus A （PVA） in GenBank to amplify the gene from the healthy plants and the plants which were infected with PVA, potato virus Y （PVY）, potato virus X （PVX） and potato virus S （PVS）, respectively. Only one PVA specific 834 bp eDNA product was amplified fi＇om the plants which were infected with PVA, but not from the others. The detectable viral dsRNA amount of this RT-PCR was 1 pg. This RT-PCR has the good sensitivity and specialty. These results indicate that this RT-PCR could be used for rapid detection of the PVA disease.