Simultaneous detection and differentiation of four viruses in passion fruit plants by a multiplex RT-PCR

Abstract A multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) assay was developed to enable the simultaneous detection and differentiation of four viruses that infect passion fruit, citrus-associated rhabdovirus (CiaRV), East Asian Passiflora virus (EAPV), latent (PLV), Telosma mosaic (TeMV). The optimized parameters included primer concentration, annealing temperature, extension time, number cycles. established RT-PCR produced corresponding products with sizes 597 bp, 529 320 235 which were specific for CiaRV, EAPV, PLV, TeMV, respectively. fragments could be distinguished clearly by agarose gel electrophoresis. limit 100 pg total RNA CiaRV 10 1.0 ng TeMV. also tested using field samples, results showed detect in single or multiple infections fruit. here will quite helpful diagnosis fruit infected various combinations field.