Random amplified polymorphic DNA analysis of Mycobacterium tuberculosis isolates resistant to Isoniazid in Indonesia

Asaad Maidin, Agnes Lidjaja, Mochammad Hatta

Biopharmaceutical Laboratory Faculty of Pharmacy Hasanuddin University, Makassar, Indonesia.
Microbiology Department Faculty of Medicine Hasanuddin University, Makassar, Indonesia.
Molecular Biology and Immunology Laboratory, Microbiology Department Faculty of Medicine Hasanuddin University, Makassar, Indonesia.

DOI: 10.4236/jtr.2013.12003                

Abstract

Background: M. tuberculosisis the most important etiological factor of tuberculosis. One of the factors that make TB hard to eradicate is the emergence of M. tuberculosisdrug resistance. Drug resistance in M. tuberculosisis attributed primarily to the accumulation of mutations in the drug target gene. Objectives: to analyze profile of Random Amplified Polymorphic DNA (RAPD) in M. tuberculosisisolates resistant to Isoniazid and found RAPD marker. Methods: seven Isoniazid resistant isolate of M. tuberculosisfrom Ma kassar, Indonesia strain were analyzed by RAPD method using primers OPN 02, OPN 09, OPN 20, BG 65, N 9, that amplification fragment DNA than as molecular marker. Results: The results of the present study showed high degree of polymerphism in theM.tuberculosisstrains in the population, and found that specific DNA fragment at Isoniazid resistant isolates using primer N 9 is 1450 bp as a marker. Conclusion: This study gives information about RAPD marker of M. tuberculosis strain to Isoniazid resistant.

Keywords

Mycobacterium tuberculosis; RAPD; Isoniazid Resistant

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Mendez, J.C. (2001) Multi drug resistance in tuberculosis and the use of PCR for defining molecular markers of resistance, Florida.
[2]	Palomino, J.C. (2006) Newer diagnostic for tuberculosis and multidrug resistant tuberculosis, mycobacteriology unit. Institute of Tropical Medicine, Antwerp.
[3]	Health Department Republic of Indonesia (2011) pedoman nasional pengendalian tuberkulosis. jakarta.
[4]	Cole, S.T., et al. (2005) Tuberculosis and the tubercle bacillus. ASM Press, Washington DC.
[5]	World Health Organization (2010) Multidrug and extensively drug-resistant TB (M/XDR-TB). Global Report on Surveillance and Response, Geneva.
[6]	Rattan, A., et al. (2006) Multidrug-resistant mycobacterium tuberculosis: Molecular perspectives. All India Institute of Medical Sciences, Ansari Nagar, New Delhi.
[7]	Newton, C.R. and Graham, A. (1994) Introduction to biotechniques PCR. Bios Scientific Publishers, Oxford.
[8]	Richner, S., Meining, J. and Kirby, R. (1999) DNA profiling of mycobacterium tuberculosis from the Eastern Cape Province of South Africa and the detection of a high level of genetic diversity, department of biochemistry and microbiology. Rhodesh University, Grahamstown.
[9]	Bardakci, F. (2001) Random Amplified Polymorphic DNA (RAPD) markers. Cumhuriyet University, Faculty of Arts and Science, Department of Biology, Tubitak.
[10]	Tazi, L., et al. (2004) Genetic diversity and population structure of mycobacterium tuberculosis in casablanca, a moroccan city with high incidence of tuberculosis. Journal of Clinical Microbiology, 42, 461-466.
[11]	Korzekwa, K., Polok, K. and Zielinski, R. (2006) Application of DNA markers to estimate genetic diversity of mycobacterium tuberculosis strains. Poland Journal of Microbiology, 55, 19-24.
[12]	Singh, J.P.N., Verma, R. and Chaudhurt, P. (2006) Random Amplified Polymorphic DNA (RAPD) analysis of mycobacterium tuberculosis strains in India. Journal of Veterinary Science, 7, 181-187.
[13]	Jain, A., et al. (2008) Prevalence of multidrug resistant mycobacterium tuberculosis in Lucknow, Uttar Pradesh. Department of Microbiology & Clinical Epidemiology Unit, CSM Medical University, Lucknow.
[14]	Va’zquez-Marrufo, G., et al. (2008) Genetic diversity among mycobacterium tuberculosis isolates from Mexican patients. Canadian Journal of Microbiology, 54, Ingenta Connect.
[15]	Rodriguez, J.C., Royo, G. and Rodriguez-Valera, F. (2009) Application of four molecular techniques for typing outbreak-associated mycobacterium tuberculosis strains. Acta Pathologica Microbiologica Et Immunologica Scandinavica.
[16]	Baron, E.J., Peterson, L.R. and Finegold, S.M. (1994) Baily & scotts diagnostic microbiology. 9th Edition, Mosby—Year Book, Toronto.
[17]	Mahon, C.R. and Manuselis, G. (1995) Textbook of diagnostic microbiology. W.B. Saunders Company, Tokyo.
[18]	Promega Corporation (2012) Wizard genomic DNA purification Kit, isolating genomic DNA from gram positive and gram negative bacteria. Madison, 16-17.
[19]	Harn, H.-J., et al. (1997) Evidence of transmission of mycobacterium tuberculosis by random amplified polymorphic DNA (RAPD) fingerprinting in Taipei City, Taiwan. Department of Pathology, General Hospital, Ting-Chow Rd, Taipei.
[20]	Linton, C.J., et al. (1995) Comparison of random amplified polymorphic DNA with restriction fragment length polymorphism as epidemiological typing methods for mycobacterium tuberculosis, Bristol.