Rapid and sensitive detection of potyvirus infecting tropical tuber crops using genus specific primers and probes

Abstract

A reverse transcription polymerase chain reaction assay using potyvirus specific primers designed from the core of the coat protein was carried out, and a cDNA fragment of 327 bp was obtained from most of the potyviruses infecting the tropical tuber crops. Reverse transcription polymerase chain reaction (RT-PCR) products were sequenced and found to be derived from the expected virus. Specific cDNA probe was generated from the amplicon, and the probe was then successfully used for the diagnosis of the potyviruses infecting the major tuber crops through biotinylated nucleic acid spot hybridisation. The specific probe developed could detect the potyviruses infecting tuber crops namely SPFMV, DsMV and DAV from sweet potato, aroids and yams respectively.

Key words: Coat protein, potyvirus, reverse transcription polymerase chain reaction, nucleic acid spot hybridisation.