Analysis of relative genes expression and mutation of pstB and efpA efflux pumps in Mycobacterium simiae isolates from suspected tuberculosis patients by using quantitative Real-time PCR.

BACKGROUND

Mycobacterium simiae is commonly found in people with weak immune systems such as the elderly and people with tuberculosis and other lung diseases. The aim of this study is the epidemiology of M. simiae infections in Iran and the world, therefore, in this study, analysis of relative gene expression and mutation of pstB and efpA efflux pumps in Mycobacterium simiae isolates from suspected tuberculosis patients by using Real-time quantitative PCR. Sixty-five sputa samples of suspected tuberculosis specimens were collected. The identification of NTM Species by PCR sequencing and determining drug sensitivity by micro broth dilution method. The investigate mutations in pstB and efpA efflux pump genes using the PCR-sequencing method. Comparative evaluation of the replication efficiency of internal control gene 16SrDNA and target genes pstB and efpA.

RESULTS

In total, 15 isolates of M. simiae were evaluated and a drug sensitivity test was performed for them against the antibiotic ethambutol, moxifloxacin, clarithromycin, and linezolid. The highest resistance to moxifloxacin and linezolid antibiotics and the lowest resistance to clarithromycin antibiotics were observed. The results of expression levels of two efflux PstB and efpA showed that there was a significant difference in the expression level of the efpA efflux pump gene in M. simiae three-resistant and double-resistant isolates compared to the sensitive group and the standard.

CONCLUSIONS

The high level of antibiotic resistance In M. simiae isolates is an alarm and monitoring all factors related to antibiotic resistance, including efflux pumps, is an important research topic.