Development of a highly sensitive multiplex reverse transcription polymerase chain reaction (m-RT-PCR) method for detection of three potato viruses in a single reaction and nested PCR [electronic resource].

Includes references.

Potato virus Y (PVY), potato virus X (PVX) and potato leaf roll virus (PLRV) are the most economically important viruses in commercial potato production in Egypt. The m-RT-PCR assays were used for the detection of these three viruses in one reaction using three specific primer pairs designed to amplify the full-length of the coat protein gene of each virus (810 bp for PVY, 700 bp for PvX, 548 bp for PLRV). Specificity of the three primer pairs was confirmed by the nested PCR (nPCR) assay using three internal specific primer pairs designed to amplify 480, 360, and 420 bp from the internal sequence of the cp gene of PVY, PYX and PLRV, respectively. The amplified products were sequenced and analyzed. This m-RT-PCR method detected all three viruses in a single reaction mixture in naturally infected field grown potatoes and nPCR assay can confirm the presence of virus infection without needing for sequence analysis. Key words: Potato, PVY, PvX, PLRV, m-RT-PCR, nPCR, sequence analysis.

Summary in Arabic.

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