DETECTION OF NARCISSUS LATENT VIRUS ISOLATES USING ONE-STEP RT-PCR ASSAY

A one-step RT-PCR procedure was developed for specific detection of Narcissus latent virus (NLV) isolates. Following alignment of RNA sequences of three NLV isolates, the conserved sequence fragments were identified in viral polyprotein gene and 3’UTR region. Based on those fragments, a forward NLs1 and two reverse: NLa1 and NLSCPR2 primers were designed. Primer pairs NLs1-NLa1 and NLs1NLSCPR2 proved to be effective in amplification of a 669 bp and 1295 bp products, respectively. Sequence analysis of amplified products confirmed their specificity.