Comparison of PCR-RFLP and Allele Specific-PCR and sequencing in rapid diagnosis of isoniazid resistance in clinical isolates of Mycobacterium tuberculosis
| Autor: | Hamid Bornasi, Mohammad Arjomandzadegan, Ahmadreza Bahrmand, Alireza Hadizadeh Tasbiti, Azam Ahmadi, Maryam Tayeboon |
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| Jazyk: | English<br />Persian |
| Rok vydání: | 2015 |
| Předmět: | |
| Zdroj: | Iranian South Medical Journal, Vol 18, Iss 2, Pp 296-303 (2015) |
| Druh dokumentu: | article |
| ISSN: | 1735-4374 1735-6954 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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