Characterization of the most common embCAB gene mutations associated with ethambutol resistance in Mycobacterium tuberculosis isolates from Iran

INTRODUCTION: Ethambutol (Emb) is one of the first-line drugs in the standard combination therapy for tuberculosis; however, due to the rapid increase in Emb resistance among clinical isolates of Mycobacterium tuberculosis (MTB), early detection of Emb resistance is desirable. As the embCAB operon is considered involved in resistance to Emb, this study aimed to analyze the most common mutations within the embCAB operon among MTB isolates from Iran to find any correlations of these mutations with Emb resistance. METHODS: A total of 307 clinical isolates of MTB were screened for Emb resistance by phenotypic drug-susceptibility testing. PCR amplification was performed on extracted DNA from all Emb-resistant and randomly selected Emb-susceptible isolates using sets of primers for various gene loci of embC, embA, and embB, followed by sequencing for the detection of most common alterations. RESULTS: In total, ten isolates showed resistance to Emb by phenotypic susceptibility testing (3.25%). The mutation rate in ten Emb-resistant MTB strains was 20% (n=2), comprising one mutation in embB (10%), at codon 306 Met–Val and one in embC (10%) at codon 270 Thr–Ile. A nonsynonymous mutation in the embA gene in one of the randomly selected Emb-susceptible isolates located in codon 330 Leu–Leu was also noticed. CONCLUSION: The majority of our Emb-resistant isolates (n=8, 80%) did not demonstrate the sequences investigated within the embCAB operon. As such, these mutations solely are insufficient for the development of complete resistance to Emb in MTB isolates. Additional mechanisms of resistance other than mutations in these sequences studied within the embCAB operon should also be considered.