Haemonchus (H.) contortus is a blood-feeding parasite causing substantial losses to sheep and goats sector across the globe. Pakistan’s livestock industry comprises of more than 100 million head of small ruminants contributing to meet the protein requirement of people. To date, there is no report on genetic characterization of H. contortus from Jhang District of Pakistan. With the aim to narrow this research gap, the current study was planned and molecular confirmation and genetic diversity of H. contortus was determined. Species-specific primers were used to amplify partial ITS-2 region. Sequencing and subsequently identity of every isolate was verified through NCBI BLAST program. Molecular analysis revealed a total of 14 haplotypes out of 30 isolates with 23 mutations. High haplotype diversity (Hd = 0.846) was observed with statistically significant Tajima’s D (-2.00336*) value suggesting population expansion. The phylogenetic analysis showed existence of unique haplotypes of H. contortus at Jhang. Further, significantly negative Tajima’s D (-2.16660*) and non-significant Fu’s Fs (-4.218) also indicated population expansion as obtained in the median-joining network consisting of a dataset of 39 sequences. The study has reflected high genetic variation and existence of unique haplotypes of H. contortus in Jhang, Pakistan. The results of this study will serve as an important database for future studies H. contortus in the country.