Virulent strains of Newcastle disease virus (NDV) can cause severe damage to tissues in the gastrointestinal tract, respiratory and reproductive organs, in addition to neurological disorders and high mortality in poultry. Many countries adapt regular vaccine application for disease control. Nevertheless, virulent NDV is still endemic, impacting the poultry industry. Accordingly, continuous virus detection and identification are required to determine the epidemiological situation of the disease and to identify the widely distributed viral strains. Additionally, genotyping of the circulating strains along with vaccination with the genotype-matched ND vaccines are crucial for disease control. Both fusion (F) and haemagglutinin– neuraminidase (HN) proteins are surface proteins that can play major roles in infection and antigenicity of NDV. The site of cleavage of fusion protein that found at positions 112 to 117 of the precursor protein, can determine viral pathogenicity. haemagglutinin– neuraminidase is a multifunctional protein that is involved in interaction with F protein to promote fusion, receptor recognition and removal, and preventing viral self-aggregation.