Multi-drug resistant gene mutation analysis in Mycobacterium tuberculosis by molecular techniques.

Autor: Sultana M; Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh., Alam MM; Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh., Mistri SK; Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh., Mostafa Kamal SM; National Tuberculosis Reference Laboratory (NTRL), Dhaka-1207, Bangladesh., Ahsan CR; Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh., Yasmin M; Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh.
Jazyk: angličtina
Zdroj: Iranian journal of microbiology [Iran J Microbiol] 2024 Aug; Vol. 16 (4), pp. 459-469.
DOI: 10.18502/ijm.v16i4.16304
Abstrakt: Background and Objectives: Rifampicin (RIF) and isoniazid (INH), two most potent antibiotics, are prescribed to cure tuberculosis. Mycobacterium tuberculosis , the causative agent of multidrug-resistant tuberculosis (MDR-TB), is resistant to these first-line drugs. Here, two molecular techniques were demonstrated such as PCR sequencing-based and GeneXpert assay for rapidly identifying MDR-TB.
Materials and Methods: Pulmonary samples (sputum) were collected from 55 MDR-TB suspected patients from the National Tuberculosis Reference Laboratory (NTRL), Dhaka where the research work was partially accomplished and continued in the department of Microbiology, University of Dhaka, Bangladesh. We strived for sequencing technique as well as GeneXpert assay to identify mutations in rpo B and kat G genes in MTB strains and sputum directly. Culture-based drug susceptibility testing (DST) was performed to measure the efficacy of the molecular methods employed.
Results: When analyzed, rpo B gene mutations at codons 531 (54.54%), 526 (14.54%), and 516 (10.91%) were found by sequencing in 80% of the samples. Nucleotide substitution at kat G315 (AGC→ACC) was spotted in 16 (76.19%) out of 21 samples. When comparing the sequencing results with DST, sensitivity and specificity were investigated to determine drug-resistance (rifampicin-resistance were 98 and 100% whereas isoniazid-resistance were 94 and 100% respectively). Additionally, as a point of comparison with DST, only 85.45% of RIF mono-resistant TB cases were accurately evaluated by the GeneXpert assay.
Conclusion: This research supports the adoption of PCR sequencing approach as an efficient tool in detecting MDR-TB, counting the higher sensitivity and specificity as well as the short period to produce the results.
(Copyright© 2024 The Authors. Published by Tehran University of Medical Sciences.)
Databáze: MEDLINE
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