The Reproducibility of the Multiplex RAPD-PCR Assay in Genotyping of Mycobacterium Tuberculosis Isolates from Sulawesi, Indonesia
American Journal of Clinical and Experimental Medicine
Volume 2, Issue 2, March 2014, Pages: 14-21
Received: Feb. 20, 2014; Published: Mar. 20, 2014
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Authors
Mochammad Hatta, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia
AndiRofian Sultan, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia
Ressy Dwiyanti, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia;Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Muhammad Sabir, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia;Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Andini Febrianty, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia;Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Ahmad Adhyka, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia;Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Nur Indah Purnamasari, Department of Microbiology, Faculty of Medicine, Haluoleo University, Kendari, Indonesia
Muhammad Reza Primaguna, Department of Microbiology, Faculty of Medicine, Haluoleo University, Kendari, Indonesia
Juhri Saning, Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Yusriani Mangarengi, Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Munawir Muhammad, Department of Microbiology, Faculty of Medicine, Tadulako University, Palu, Indonesia
Nataniel Tandirogang, Department Microbiology, Faculty of Medicine, Mulawarman University, Samarinda, Indonesia
Yadi Yasir, Department Microbiology, Faculty of Medicine, Mulawarman University, Samarinda, Indonesia
Masyhudi Amir, Department Microbiology, Faculty of Medicine, Mulawarman University, Samarinda, Indonesia
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Abstract
Background: Random Amplified Polymorphic DNA (RAPD) assay has recently emerged as a genotyping method which is both robust and highly discriminatory for bacterial strain differentiation. However, RAPD assessment for Mycobacterium tuberculosiscomplex (MTC) isolates is still limited. Despite its simplicity and rapidity, conventional RAPD also has low reproducibility due to its sensitivity to several factors. Therefore we studied the feasibility of an RAPD-PCR assay to define the genetic diversity of MTC isolates and to evaluate its reproducibility.Methods:493clinical MTC isolates from the island of Sulawesi in Eastern Indonesia, collected from 2005-2012were subjected to Multiplex RAPD assay using 11 random decamer primers instead of one primer which is common in conventional RAPD. All 11 primers were found to be differentiated and produced specific RAPD profiles. The polymorphic amplicons served as RAPD markers for MTC. The dendrograms, obtained by different primers, showed the discriminatory ability of the primers.Results:Multiplex RAPD-PCR results show that the majority of the isolates from South Sulawesi, Southeast Sulawesi andCentral Sulawesiin eastern region of Indonesia belong to group MT-C (80.7%, 80.0% and 62.6 % respectively) with result reproducibility as high as 100%. Conclusion:Molecular typing with multiplex RAPD-PCR is a powerful approach to show the genetic heterogeneity of MTC isolates. The discrimination power and reproducibility of this multiple loci-based RAPD was higher than conventional fewer loci-targeted RAPD.
Keywords
RAPD-PCR, Genotyping, Mycobacterium Tuberculosis Complex, Sulawesi Indonesia
To cite this article
Mochammad Hatta, AndiRofian Sultan, Ressy Dwiyanti, Muhammad Sabir, Andini Febrianty, Ahmad Adhyka, Nur Indah Purnamasari, Muhammad Reza Primaguna, Juhri Saning, Yusriani Mangarengi, Munawir Muhammad, Nataniel Tandirogang, Yadi Yasir, Masyhudi Amir, The Reproducibility of the Multiplex RAPD-PCR Assay in Genotyping of Mycobacterium Tuberculosis Isolates from Sulawesi, Indonesia, American Journal of Clinical and Experimental Medicine. Vol. 2, No. 2, 2014, pp. 14-21. doi: 10.11648/j.ajcem.20140202.12
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