The aim of this study was to investigate the significance of multiple-mutations in the katG gene, predominant
nucleotide changes and its correlation with high level of resistance to
isoniazid in Mycobacterium tuberculosis isolates that were randomly collected from sputa of 42 patients with primary and secondary active
pulmonary tuberculosis from different geographic regions of Iran.
Drug susceptibility testing was determined using the CDC standard conventional proportional method.
DNA extraction, katG gene amplification, and
DNA sequencing analysis were performed. Thirty four (80%) isolates were found to have multiple-mutations (composed of 2-5 mutations) in the katG gene. Increased number of predominant mutations and
nucleotide changes were demonstrated in
codons 315 (AGC-->ACC), 316 (GGC-->AGC), 309 (GGT-->GTT) with a higher frequency among patients bearing secondary
tuberculosis infection with elevated levels of resistance to
isoniazid (MIC ≥ 5-10 µg/mL). Furthermore, it was demonstrated that the combination of mutations with their predominant
nucleotide changes were also observed in
codons 315, 316, and 309 indicating higher frequencies of mutations among patients with
secondary infection respectively. In this study, 62% (n= 21) of multi-mutated isolates found to have combination of mutations with predominant
nucleotide changes in
codons 315 (AGC-->ACC), 316 (GGC-->GTT), 309 (GGT-->GGT), and also demonstrated to be more frequent in isolates of patients with
secondary infections, bearing higher level of resistance to
isoniazid (≥ 5-10 µg/mL).