Analysis of the resistance of M. tuberculosis to fluoroquinolon and the implementation of nuclear based biomolecular technique.

Mukh Syaifudin, Dewi Septiani

Abstract


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


Full Text:

Untitled

References


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

Austria.

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,

, S77-82.

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.




DOI: http://dx.doi.org/10.14499/indonesianjpharm0iss0pp120-127

Refbacks

  • There are currently no refbacks.




Copyright (c) 2017 INDONESIAN JOURNAL OF PHARMACY

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Indonesian J Pharm indexed by:

                                               

web
analytics View My Stats