The study investigated Newcastle Disease Virus (NDV) distribution across different regions of Pakistan's Khyber Pakhtunkhwa province. A total of 400 tissue samples, i-e, proventriculus, liver, lungs, trachea (dead birds), and oropharyngeal, cloaca swabs (live birds) were collected from Peshawar, Hazara, Southern, and Malakand, revealing varied NDV positivity rates. Malakand had the highest rate (25.7%), followed by Peshawar and Hazara (20%), and the Southern region (17.8%). Statistical analysis indicated potential significant differences in NDV prevalence among regions (P-value = 0.031, Fisher's exact test). Among the 400 samples, 80 underwent RNA extraction and cDNA synthesis. PCR amplification of the F gene showed successful results in 62 samples, indicating mesogenic and velogenic strains. Further assays categorized 34 samples as velogenic/mesogenic, 20 as lentogenic, and 28 as mixed strains. Among 32 positive velogenic/mesogenic samples, 25 remained undigested with the BglI enzyme, confirming their velogenic nature due to the presence of the fusion protein cleavage site. The current study shows NDV's regional distribution, revealing their diversity in the KP Province of Pakistan. These findings are useful for better management of NDV in the future, offering insights for potential control measures against this infectious disease.