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Rapid detection of Mycobacterium tuberculosis complex by loop mediated isothermal amplification
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Elham Kohan   1, Mohammad Hassan Shahhosseiny2 , Mohammad Rezd Razavi3 , Kazem Parivar4 , Elham Moslemi5 |
1- PhD Student of Cell and Molecular Biology, Department of Biology, Member of Scientific Board, Islamic Azad University, Arsanjan Branch, Arsanjan, Iran , Kohan@iaua.ac.ir
2- PhD of Biologic Products, Department of Microbiology, Islamic Azad University, Qods Branch, Tehran, Iran
3- PhD of Molecular Biology, Department of Mycobacteriology, Pasteur Institute Of Iran, Tehran, Iran
4- PhD of Developmental Cellular Biology, Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Science and Research Campus, Tehran, Iran
5- PhD of Cellular and Molecular Biology, Department of Biology, Islamic Azad University, Qiamdasht Branch, Qiamdasht, Iran |
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Abstract: (14573 Views) |
Background: Rapid and efficient diagnosis is one of the crucial issues in the control of tuberculosis. Loop mediated isothermal amplification (LAMP) is a simple and rapid technique for nucleic acid amplification. In this study, we evaluate a LAMP assay targeting IS6110 gene for diagnosis of M. tuberculosis in clinical samples.
Materials and Methods: 155 clinical sputum specimens were collected from suspected patients with tuberculosis. Six specific primers were designed for LAMP assay. LAMP reaction was performed under optimal condition. Sensitivity and specificity of LAMP were determined.
Results: A successful LAMP reaction with newly designed primers was carried at 66ºC for 60 minutes. The detection limit of LAMP was 5fg DNA per tube. The sensitivity and specificity of this assay in clinical sputum samples were 100% and 95.9%, respectively.
Conclusion: LAMP is a rapid, sensitive and low cost technique that can be suitable for routine usage in diagnosis of TB. |
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| Keywords: Mycobacterium tuberculosis complex, Rapid detection, LAMP technique, PCR |
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Full-Text [PDF 197 kb]
(3215 Downloads)
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Semi-pilot: Experimental |
Subject:
Molecular Biology
Received: 2012/06/27 | Published: 2012/06/15
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