RAPID DETECTION OF SUSCEPTIBILITY TO ETHAMBUTOL IN CLINICAL MYCOBACTERIUM TUBERCULOSIS ISOLATED FROM TUBERCULOSIS PATIENTS BY PCR- RFLP

Eethambutol is a key drug to treatment of tuberculosis; and resistance against it
has been increasingly reported. In this study, Polymerase Chain Reaction -Restriction Fragment
Length Polymorphism (PCR-RFLP) method in rapid detection of tuberculoses mycobacterium strains
against the ethambutol has been investigated.
Materials & Methods: This study was conducted on 60 strains that were selected from 127 strains in
Tuberculosis and Pediatric Infections Research Center. DNA was extracted by Chelex 100 method.
PCR test was performed using specific primers for embB gene. Digestions of PCR products with
HaeIII and NlaII restriction enzymes, then the pattern of restriction fragments generated were
analyzed. The section was sequenced in a few samples, and it was compared to the presented method
as golden standard.
Results: Out of 60 studied stains, 43 were phenotypically resistant to ethambutol, and 17 were
sensitive. PCR revealed that the band 167 showed the correct selection of primers and the appropriate
plan of amplification. Out of 43 resistant strains, 19 stains were diagnosed to have mutation in ATGMet
codon 360 using RFLP method, and 24 were found to be non-mutant.
Conclusion: According to the findings, that PCR-RFLP can be a simple and rapid method to diagnose
the ethambutol -sensitivity in tuberculosis mycobacterium strains.