Quick Detection of Potato Viruses by Duplex RT-PCR

In order to establish a technique using RT-PCR to detect potato viruses,which was rapid,accurate and sensitive,the primers for PVX,PVS,PVA and PLRV fragments were designed based on their coat protein,and a duplex reverse transcription polymerase chain reaction(d-RT-PCR) was optimized from reverse transcription(RT) stage and polymerase chainreaction(PCR) stage.The optimized d-RT-PCR can amplify PVX,PVS,PVA and PLRV simultaneously,and the fragments were 620 bp(PVX),435 bp(PVS),300 bp(PVA), 222bp(PLRV).The result showed that antisense primer rations of two viruses,dNTPs concentration and Mg2+ concentration had great impacts on the detection.The duplex RT-PCR was also suitable for viruses detection of propotato seed both from tissue cultural factory and from naturally field growing potato plant.