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:: Volume 18, Issue 3 (Iranian South Medical Journal 2015) ::
Iran South Med J 2015, 18(3): 547-555 Back to browse issues page
Simultaneous detection of TB and drug resistance to Isoniazid in Mycobacterium tuberculosis clinical isolates using PCR-RFLP method
Maryam alsadat Tayeboon1, Maryam Sadrnia2, Hamidreza Mohajerani3
1- Infectious Diseases Research Center, Arak University of Medical Sciences, Arak, Iran
2- Department of Biology, Payame Noor University, I.R. of Iran , msadrnia@yahoo.com
3- Department of Microbiology, Islamic Azad University, Arak Branch, Arak, Iran
Abstract:   (5034 Views)

Background: Early detection and suitable drug admission are the first attempts to control of TB and MDR-TB. In this study, PCR-RFLP method was used for rapid detection of Mycobacterium tuberculosis and resistance to isoniazid. Materials and Methods: In the present study, 87clinical isolates of MTB were used in a PCR-RFLP method. The RFLP-PCR assay by specific primer was used for two purposes In the first, PCR of katG gene for diagnosis of bacteria and in the second step, PCR product by endonuclease enzyme in a RFLP reaction produced a pattern in electrophoresis for mutation detection in katG315. Sequencing method was used for confirmation of RFLP results. Results: The study showed that all studied strains produced band 620bp that confirmed MTB. From the 46 resistant strains to isoniazid, the RFLP-PCR method showed that 44 strains had mutations in the katG gene Ser315Thr. The 41 susceptible strains had not any mutation at the codon. Results of sequencing were confirmed results of the molecular method. Sensitivity of RFLP-PCR test was 95.6% (95% CI :0.85-0 .98) and specificity was 100% (95% CI :0.91-1 .0). Conclusion: RFLP-PCR test is a good tool for simultaneous diagnosis and detection of katG315 mutation in clinical isolates of MTB.

Keywords: Mycobacterium tuberculosis, diagnosis, drug resistance, katG, PCR-RFLP
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Type of Study: Original | Subject: Communicable Diseases
Received: 2013/05/12 | Accepted: 2014/05/17 | Published: 2015/06/28
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Tayeboon M A, Sadrnia M, Mohajerani H. Simultaneous detection of TB and drug resistance to Isoniazid in Mycobacterium tuberculosis clinical isolates using PCR-RFLP method. Iran South Med J. 2015; 18 (3) :547-555
URL: http://ismj.bpums.ac.ir/article-1-701-en.html


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Volume 18, Issue 3 (Iranian South Medical Journal 2015) Back to browse issues page
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