Detection and differentiation of pigeon PMV-1 by restriction site analysis of F gene region

Detection and pathotyping of Newcastle disease virus (NDV) are extremely important because the appearance of virulent virus has significant economic consequences. Pigeon Paramyxovirus serotype-1 (PPMV-1) present difficulty when assessed by conventional in-vivo pathogenicity tests. The technique of RT-PCR and restriction enzyme analysis was used to detect and differentiate PPMV-1. Digestion of RT-PCR amplified F gene sequences, encoding for the cleavage activation sites of fusion protein was carried out with restriction enzymes BglI, HhaI, HaeIII, HinfI, MboI, MspI, PvuII and StyI. Restriction enzyme digestion of the amplicon by BglI, HhaI, MboI and either MspI or HaeIII could differentiate PPMV-1 isolates from reference lentogenic, mesogenic and viscerotropic velogenic NDV strains. Restriction enzyme HinfI and not BglI could differentiate between virulent and non-virulent NDVs. Based on restriction profile with enzymes StyI, PvuII and HhaI, seven PPMV-1 isolates in India could be divided into four distinct groups.