Quick Detection of Multiple Potato Viruses by Multiplex RT-PCR

The primers for potato virus X(PVX),potato virus S(PVS),potato virus A(PVA) and potato leaf roll virus(PLRV) fragments were designed based on their coat protein region,and a multiplex reverse transcription polymerase chain reaction(m-RT-PCR) was optimized from dNTPs concentration,annealing temperature,Mg2+ concentration and cyclic conditions.The optimized m-RT-PCR can amplify PVX,PVS,PVA and PLRV simultaneously,and the fragments were 620 bp(PVX),435 bp(PVS),300 bp(PVA) and 222 bp(PLRV).The results showed that the dNTPs concentration at reverse transcription stage(RT) and the Mg2 + concentration at PCR stage had a great impact on the detection performance,and next was the annealing temperature at PCR stage;the cyclic conditions has small impact on the detection results.The optimized m-RT-PCR provide a quick,sensitive and simple method for detecting potato viruses,and it also provide a effective way for early detection and control of potato viruses