The effect of anti-tuberculosis drugs therapy on mRNA efflux pump gene expression of Rv1250 in Mycobacterium tuberculosis collected from tuberculosis patients.

Abstract

Efflux pumps are transmembrane proteins that vigorously participate in extruding a wide range of substrates, including drugs, outside the bacterial cell. We aimed to investigate the mRNA expression level of the Rv1250 efflux pump gene in Mycobacterium tuberculosis isolated from individuals with tuberculosis who received drug therapy, at the 1st, 3rd and 5th months, and newly diagnosed patients with tuberculosis who will receive drug therapy (0 month). The study was a multiple cross-sectional longitudinal design—50 different M. tuberculosis isolates and a reference strain H37Rv were subcultured in LJ medium and confirmed by multiplex PCR for identification of M. tuberculosis and collected for RNA extraction. Total bacterial mRNA was analysed using real-time quantitative PCR to evaluate mRNA quantification gene expression. There were differences in the level of Rv1250 mRNA expression between sensitive (n = 11) and resistant (n = 40) groups of 5.961 ± 0.414 and 10.192 ± 1.978, respectively (fold changes; p < 0.05). There were significant differences of expression level among M. tuberculosis-resistant groups (p < 0.05) specifically 7.573 ± 0.424 for 0-month drug therapy (n = 10), 9.438 ± 0.644 for 1st month drug therapy (n = 10), 11.057 ± 0.262 for 3rd month drug therapy (n = 10) and 12.701 ± 0.460 for 5th month drug therapy (n = 10). We assume that the extent of Rv1250 gene expression in M. tuberculosis clinical isolates is a result of the exposure to antimicrobials during treatment, which affect the basic expression of the efflux pump Rv1250 gene.