:: Volume 18, Issue 2 (Iranian South Medical Journal 2015) ::
Iran South Med J 2015, 18(2): 296-303 Back to browse issues page
Comparison of PCR-RFLP and Allele Specific-PCR and sequencing in rapid diagnosis of isoniazid resistance in clinical isolates of Mycobacterium tuberculosis
Hamid Bornasi1, Mohammad Arjomandzadegan2, Ahmadreza Bahrmand3, Alireza Hadizadeh Tasbiti3, Azam Ahmadi4, Maryam Tayeboon4
1- Department of Immunology and Microbiology, Arak University of Medical Sciences, Arak, Iran
2- Department of Immunology and Microbiology, Arak University of Medical Sciences, Arak, Iran
Tuberculosis and Pediatric Infectious Diseases Research Center, Arak University of Medical Sciences, Arak, Iran , arjomandzadegan@arakmu.ac.ir
3- Mycobacteriology Dept, Pasteur Institute of Iran, Tehran
4- Tuberculosis and Pediatric Infectious Diseases Research Center, Arak University of Medical Sciences, Arak, Iran
Abstract:   (4996 Views)

Background: Isoniazid is from the first-line drugs for treatment of tuberculosis. Resistance to Isoniazid is increasingly reported. In the study, molecular methods for rapid detection of isoniazid resistance in clinical isolates of Mycobacterium tuberculosis were compared. Materials and Methods : Sixty clinical isolates of Mycobacterium tuberculosis were selected and their resistance and susceptibility were determined by proportional method. Molecular methods of PCR-RFLP and Allele Specific-PCR (AS-PCR) for determination of probably mutation in codon 315 of katG gene that related to Isoniazid resistance were used and compared by sequencing results as gold standard. In AS-PCR specific primers by proper PCR conditions were used. RFLP analysis of the PCR products was performed using the enzyme HpaII. Results: From 60 studied strains, 27 isolates were resistant and 33 strains were susceptible to isoniazid. Sequencing results showed that AS-PCR and PCR-RFLP well able to detect mutations in codon 315 of the strains by 81.48% sensitivity and 100% specificity respectively. None of the susceptible strains harbored mutation in katG315. Conclusion: Our results indicated that PCR-RFLP and AS - PCR methods were simple and fast methods for determination of isoniazid resistance in Mycobacterium tuberculosis. These rapid and low cost methods is recommended simultaneously to reduce the risk of error in routine work.

Keywords: Isoniazid, Mycobacterium tuberculosis, PCR-RFLP, Allele Specific-PCR
Full-Text [PDF 647 kb]   (2042 Downloads)    
Type of Study: Original | Subject: General
Received: 2013/12/5 | Accepted: 2014/02/1 | Published: 2015/05/3


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