Newcastle disease (ND) which is caused by Avian Paramyxovirus type I is among the major concerns of poultry production worldwide. In developing countries especially where backyard poultry are not monitored for infection, ND remains a threat. The use of vaccines (live and inactivated) for the last 50 years in South Asia has not been able to control the infection. The poultry mortality rate worsened after the emergence of Avian Influenza in commercial poultry during early 1990's. The present study was conducted on 84 samples during 19952069, from commercial poultry flocks in the suburbs of Karachi to isolate and characterize strains of Newcastle disease virus circulating in the region. The samples were passaged in 9-10 day old chicken embryonated eggs. Haemagglutinating activity was observed in eighty samples which were cross checked using ND virus (NDV) polyclonal serum employed in haemagglutination inhibition assay and only forty seven were positive for NDV. The infective allantoic fluid was later used for pathotyping of isolates by employing biological characterization tools. Results demonstrated that the majority of isolates were either mesogenic or velogenic strains based on the mean death time and intra cerebral pathogenicity indices. Molecular identification was done using reverse transcription polymerase chain reaction (RT-PCR) performed using viral RNA extracted from infective allantoic fluid. Three sets of previously reported primers were employed and primer set BK1/BK2 was found to be the most sensitive. The results suggest that Newcastle disease remains endemic and requires similar attention to that given to highly pathogenic avian influenza.