Development of one-step multiplex RT-PCR for simultaneous detection of four potato viruses

Abstract:

One-step multiplex reverse transcription polymerase chain reaction (M-RT-PCR) was established and optimized for the simultaneous detection of potato virus X, potato virus Y, potato virus A and potato leafroll virus. The primers for PVX (732 bp), PVY (422 bp), PVA (132 bp) and PLRV (336 bp) fragments were designed based on viral coat protein gene. The sensitivity of detection was about 7.8 pg/μL RNA. No fragments were obtained from PVM, PVS, AMV, TMV and PSTVd. The results showed that it was a specific, sensitive detecting method and quicker, simpler, more efficient and cheaper than two-step multiple RT-PCR. It was provided an effective method of the detection for potato virus.

Key words:One-step;Multiplex reverse transcription and polymerase chain reaction (M-RT-PCR);Virus detection;Potato