Evaluation of Multiplex Reverse Transcription Polymerase Chain Reaction (RT-PCR) for Simultaneous Detection of Potato Viruses and Strains

Abstract

A multiplex reverse transcription polymerase chain reaction (RT-PCR) protocol was evaluated for the

simultaneous detection of <i>Potato leaf roll virus </i>(PLRV), <i>Potato virus </i>X (PVX) and <i>Potato Virus Y </i>(PVY). The

multiplex RT-PCR detection of viruses was equally efficient whether RNAs were extracted using commercial kits or

a sodium sulphite based nucleic acid extraction procedure. cDNAs were prepared either using a common primer

(oligo dT) or specific antisense primer followed by specific primer pairs for PCR. The multiplex RT-PCR separation

of strains of PVY was accomplished by using a competitive multiplex RT-PCR (with one antisense and two sense

primers). The multi virus or multistrain detection approaches described here have potential application to other

crop-virus combinations.