This study aimed to identify the spectrum of mutations contributing to first-line drug resistance in MDR-TB isolates from Khyber Pakhtunkhwa, Pakistan.<h4>Methods</h4>Whole-genome sequencing was performed on 16 clinical isolates (12 MDR and 4 drug-susceptible) to identify resistance-associated mutations in <i>rpoB</i>, <i>inhA</i>, <i>katG</i>, <i>embA</i>, <i>embB</i>, <i>embC</i>, and <i>pncA</i>. This evidence concerns the gene INHA and tuberculosis.