طراحي و بررسي مقايسه اي روشهاي مولكولي تشخيص سريع مقاومت به داروهاي تزريقي ضد سل در سويه هاي كلينيكي مقاوم به داروهاي تزريقي

عنوان انگليسی

Designing and Comparison Study of Rapid Detection Methods of Resistance to Injectable Drugs in Clinical Strains of Mycobacterium tuberculosis

خلاصه انگلیسی

Background & Objectives: In this study some molecular methods were designed for rapid detection of resistance to kanamycin and amikacin. Methods: Among 120 clinical isolates of Mycobacterium tuberculosis, 52 strains were selected for evaluation of possible mutations. A PCR-RFLP method was designed for detection of wild type (using enzyme AjiI) and mutant form (enzyme BstFNI) of the isolates. Furthermore, Multiplex Allele Specific Method (MAS PCR) was design for detection mutations in codons 1401 and 1402 gene rrs. Some selected isolates were sequenced. Results: Among the 48 strains were examined by PCR-RFLP, BstFNI enzyme could detect 15 mutant strains among 18 phenotypically resistant and 29 non-mutant isolates from 30 susceptible isolates. The sensitivity of this method was 83.33% and specificity was 96.66%. In the other hand, 12 mutants from 20 resistant strains and 29 non-mutant strains from 32 susceptible strains were detected by AjiI enzyme. The sensitivity and specificity of this method was 60% and 90.62%, respectively. In MAS PCR, 3 mutants from 6 resistant strains and 12 non-mutants from 17 resistant strains were detected. The sensitivity of this method was 50% and specificity was 70.58%. Results of sequencing method were proved the results of molecular methods. Conclusion: PCR-RFLP method by BstFNI enzyme was the best method for rapid detection of Mycobacterium tuberculosis resistant to second-line injectable drugs and was recommended for routine use.

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