Analysis of the resistance of M. tuberculosis to fluoroquinolon and the implementation of nuclear based biomolecular technique.
Tuberculosis (TB) is still a problem in community health with high rate of mortality. The case became much more complicated due to emerge of Mycobacterium tuberculosis which are resistant to the drugs. This caused the movement of attention from the first line drugs to fluoro-quinolon (FQ) as alternative drug. The aim of this research was to do analysis the mutation which causing the resistance of bacterial through nucleic acid alterations with polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) technique. Analysis was done on gyrA and gyrB genes encoding DNA gyrase of bacterial and closely related to FQ resistance in 100 of sputa samples of positive BTA test results. DNA of M. tuberculosis strain H37Rv was used as control. From analysis on gyrA gene it was known that 57 samples were positive PCR and no resistant sample was found. For gyrB gene, only 12 of them were positive PCR and again there was no samples had mutation as cause of resistance. These mean that FQ could be used as replacement drug. Molecular detection technique was known fast and specific for assessing bacterial resistance. Researcher proves that searching for P32-radioisotope labeled DNA alteration was more sensitive. Hopefully this results of experiment can be implemented in medication with more effective and support diagnose results so that it will lowering the risk of patient mortality.
Key words : M. tuberculosis, fluoro-quinolon, resistance, PCR, SSCP
Chaubert, P., Bastita, D. and Behattar, J., 1993, An improved method for rapid screening of DNA mutations by nonradioactive single-strand conformation polymorphism procedure, BioTechniques, 15, 586.
Cohn, D., Bustreo, F., and Raviglione, M., 1997, Drug-Resistant Tuberculosis: Review of the Worldwide Situation and the WHO/IUATLD Global Surveillance Project, Clinical Infectious Diseases, 24 (Supplement 1), S121-130.
Davis, L.G., Kuehl, W.M., and Battey, J.F., 1994, Basic Methods in Molecular Biology, Edisi kedua, Appleton and Lange, USA.
Department of Health, Republic of Indonesia, 1999, Proposed national health research priorities: the view of National Institute of Health Research and Development (NIHRD). Jakarta, Indonesia. Ministry of Health Republic of Indonesia.
Drosten, C., Panning, M. and Kramme, S., 2003, Detection of Mycobacterium tuberculosisby real-time PCR using Pan-Mycobacterial primers and a pair of fluorescence resonance energy transfer probes specific for the M. tuberculosiscomplex, Clinical Chemistry, 49, 1659-1661.
Ellison, J., Dean, M. and Goldman, D., 1993, Efficacy of fluorescence-based PCR-SSCP for detection of point mutations, Biotechniques, 15, 684-691.
Farmer, P., 1999, Immodest claims of causality In: Infections and Inequalites: The Modern Plagues, University of California Press, Berkeley, CA; pp. 231-261.
Garcia, L., Alonso-Sanz, M., Rebollo, M.J., Tercero, J.C., and Chaves, F., 2001, Mutations in the rpoB gene of rifampin-resistant Mycobacterium tuberculosis isolates in Spain and their rapid detection by PCR-Enzyme-Linked Immunosorbent Assay, Journal of Clinical Microbiology, 39(5), 1813-1818.
Hayashi, K. and Yandell, D.W., 1993, How sensitive is PCR-SSCP?, Human Mutation, 2, 404-414.
Hayashi, K., 1992, PCR-SSCP: a method for detectionof mutations, Genet. Anal. Biomol. E., 9, 73-79.
Higgins, N.P., Peebles, C.L., Sugino, A., and Cozzarelli, N.R., 1978, Purification of the subunits of Escherichia coli DNA gyrase and reconstitution of enzyme activity, Proc. Natl. Acad. Sci. USA, 75, 1773-1777.
International Atomic Energy Agency, 2000, Combating infection in developing countries, Vienna
Iseman, M.D. and Madsen, L.A., 1992, Drug-resistanttuberculosis, Clin. Chest. Med., 10, 341-353.
Jenkins, F.J., 1994, Basic methods for the detection of PCR products, PCR Methods and Applications,
Karyadi, E., Schultink, W., Nelwan, R.H.H., Gross, R., Amin, Z., Dolmans, W.M.V., Schlebusch, H., and van der Meer, J. W. M., 2000, Poor Micronutrient Status of Active Pulmonary Tuberculosis Patients in Indonesia, Journal of Nutrition, 130, 2953-2958.
Kim, B.J., Lee, K.H., Park, B.N., Kim, S.J., Bai, G.H., Kim, S.J., and Kook, Y.H., 2001, Differentiation of Mycobacterial species by PCR-restriction analysis of DNA (342 base pairs) of the RNA polymerase gene (rpoB), J. Clinical Microbiology, 39(6), 2102-2109.
Kirschner, P., Springer, B., Vogel, U., Meier, A., Wrede, A., Kiekenbeck, M., Bange, F.C., and Böttger, E.C., 1993, Genotypic identification of mycobacteria by nucleic acid sequence determination: report of a 2-year experience in a clinical laboratory, Journal of Clinical Microbiology, 31, 2882-2889.
Moore, D.F. and Curry, J.I., 1998, Detection and identification of Mycobacterium tuberculosisdirectly from sputum sediments by Ligase Chain Reaction, J. Clin. Microbiol., 36(4), 1028–1031.
Musser, J., 1995, Antimicrobial Agent Resistance inmycobacteria: molecular genetic insights, Clin. Microbiol. Rev., 8, 496-514.
Piatek, A.S., Telenti, A., Murray, M.R., El-Hajj, H., Jacobs, W.R. Jr., Kramer, F.R., and Alland, D., 2000, Genotypic Analysis of Mycobacterium tuberculosis in Two Distinct Populations Using Molecular Beacons: Implications for Rapid Susceptibility Testing, Antimicrobial Agents and Chemotherapy, 44(1), 103-110.
Ramaswamy, S. and Musser, J., 1998, Molecular genetic basis of antimicrobial agent resistance in Mycobacterium tuberculosis: 1998 Update, Tubercle and Lung Disease, 79(1), 3-29.
Rattan, A., Kalia, A., and Ahmad, N., 1998, Multidrug-resistant Mycobacterium tuberculosis: molecular perspectives, Emerging Infectious Diseases, 4(2), 195-210.
Raviglione, M., Sinder, D., and Kochi, A., 1995, Global epidemiology of tuberculosis-morbidity and mortality of a worldwide epidemic, JAMA, 273, 220-226.
Rogers, A., 1979, Techniques of autoradiography, 3rd Edition. Elsevier, North Holland, p.429.
Takiff, A.E., Salazar, L., Guerrero, C., Philipp, W., Huang, W.M., Kreiswirth, B., Cole, S.T., Jacobs,W.R., Jr, and Telenti, A., 1994, Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations, Antimicrobial Agents and Chemotherapy, 8, 77-80.
Telenti, A., Imboden, P., Marchesi, F., Lowrie, D.,Cole, S., Colston, M.J., Matter, L., Schopfer, K.,and Bodmer, T., 1993, Detection of rifampin-resistance mutations in Mycobacterium tuberculosis, Lancet, 341, 647-650.
Unniraman, S., Chatterji, M. and Nagaraja, V., 2002, DNA Gyrase genes in Mycobacterium tuberulosis: a Single Operon Driven by Multiple Promoters, J. of Bacteriology, 184, 5449-5456.
World Health Organization, 2000, World TB Day 2000: Forging New Partnerships to Stop TB, Jenewa.
World Health Organizations, 2003, PPM DOTS in Indonesia; A strategy for action. Geneva, Switzerland.
- There are currently no refbacks.
Copyright (c) 2017 INDONESIAN JOURNAL OF PHARMACY
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Indonesian J Pharm indexed by:View My Stats