Newcastle disease (ND) is an economically important, contagious poultry viral disease reported across the globe. No recent reports on ND circulating in Malaysia. Therefore, the aim of the study is to characterize 16 Newcastle disease viruses (NDVs) isolated from chickens in Malaysia in the year of 2019. All isolates were genotypically analyzed using reverse transcription-polymerase chain reaction (RT-PCR) with primers specific to the viral fusion (F) protein gene. Analysis of the F protein cleavage site’s deduced amino acid sequences revealed that from the Newcastle disease virus (NDV) isolates, three of them were virulent with two different motifs of 112RRQKRF117 and 12RRRKRF117 while other isolates were avirulent. Phylogenetic analysis demonstrated that three isolates were grouped in genotype VII, five in genotype I while eight in genotype II. All genotype VII isolates were clustered under sub-genotype VII.2 (VIIh and VIIi) which is the same strain causing previous outbreaks in Malaysia. Therefore, findings in this study demonstrated that there is no new introduction of NDV genotypes in Malaysia. However, farms should implement biosecurity measures at strict level as well as executing continuous monitoring and surveillance of the disease as these implementations would help them to conduct proper preventive measures and control of panzootic viruses in future.