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Molecular analysis of multidrug resistantMycobacterium tuberculosis isolates from Morocco

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Abstract

Tuberculosis remains a global threat to public health. Considerable efforts have been made to combat this disease. However, the emergence ofMycobacterium tuberculosis (Mtb) strains resistant to the major anti-tuberculosis drugs especially multidrug resistant (MDR) strains poses a deadly threat to control programs. The present study aims to identify the most common mutations within multidrug-resistantM. tuberculosis Moroccan isolates in order to use them as molecular markers for early and rapid detection of multidrug resistant strains. For that, allM. tuberculosis isolates received during 2002–2003 in the National Reference Laboratory of Tuberculosis in Morocco were subject to drug susceptibility tests for rifampicin and isoniazid and to a PCR probe method to detect specific mutation. Sequencing was performed for all genotypic rifampicin resistant isolates and also for four genotypic iso niazid resistant asolates randomly selected. Out of 187M. tuberculosis positive cultures, 46 (24.6%) were phenotypically resistant to both rifampicin and isoniazid. Nucleotide mutations in rpoB531, rpoB526, rpoB516, katG315 and inh-15 codons associated with resistance to rifampin (RIF) and isoniazid (INH) were found respectively in 37/46 (80.4%) and 43/46 (93.5%) isolates. Genotypic multi drug resistance was then confirmed in 74% (34/46) isolates. The mutations at codon 315 ofkatG gene and at codons 531, 526 and 516 ofrpoB gene are frequently found in MDR isolates which confirm their strong implication in the development of multidrug resistant tuberculosis. We concluded that these mutations are useful as molecular markers for detection of multidrug resistant isolates but are not yet sufficient to fully predictM. tuberculosis multidrug resistance.

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References

  • Acar J.F., Goldstein F.W. (1998). Consequence of increasing resistance to antimicrobial agents. Clin. Infect. Dis., 27 Suppl 1: S125–130.

    Article  Google Scholar 

  • Brown T.J., Tansel O., French G.L. (2000). Simultaneous identification and typing of multidrug-resistantMycobacterium tuberculosis isolates by analysis of pncA and rpoB. J. Med. Microbiol., 49: 651–656.

    CAS  PubMed  Google Scholar 

  • Cardoso R.F., Cooksey R.C., Morlock G.P., Barco P., Cecon L., Forestiero F., Leite C.Q., Sato D.N., Shikama Mde L., Mamizuka E.M., Hirata R.D., Hirata M.H. (2004). Screening and characterisation of mutation in isoniazid-resistantMycobacterium tuberculosis isolates obtained in Brazil. Antimicrob. Agents Ch., 48 (9): 3373–3381.

    Article  CAS  Google Scholar 

  • Carpel G., Fissette K., Limbana V., Van Deun A., Vandenbulcke W., Portaels F. (1995). Drug resistant tuberculosis in sub-sahara Africa: an estimation of incidence and cost for the year 2000. Tubercle Lung Dis., 76: 480–486.

    Article  Google Scholar 

  • Cavusoglu C., Hilmioglu S., Guneri S., Bilgic A. (2002). Characterization of rpoB mutations in rifampin-resistant clinical isolates ofMycobacterium tuberculosis from Turkey by DNA sequencing and line probe assay. J. Clin. Microbiol., 40: 4435–4438.

    Article  CAS  PubMed  Google Scholar 

  • Cockerill F.R. 3rd (1999). Genetic methods for assessing antimicrobial resistance. Antimicrob. Agents Ch., 43 (2): 199–212.

    CAS  Google Scholar 

  • David H., Levy Frebault V., Thorel M.F. (1987). Methodes de Laboratoire pour Mycobactériologie Clinique. Unité de tuberculose et des mycobacteries, Institut Pasteur Paris.

    Google Scholar 

  • El Messaoudi M.D., Chetioui F., Messaoudi M., Boulahbal F., Bencheikh N. (2000). Etude de la résistance duMycobacterium tuberculosis aux antibacillaires à la wilaya de Casablanca (Maroc). WHO/IUATLD Global project on Anti-Tuberculosis Drug Resistance Surveillance, WHO/CDS/TB/2000.278.

  • Fang Z., Doig C., Rayner A., Kenna D.T., Watt B., Forbes K.J. (1999). Molecular evidence for heterogeneity of the multiple-drug-resistantMycobacterium tuberculosis population in Scotland (1990 to 1997). J. Clin. Microbiol., 37 (4): 998–1003.

    CAS  PubMed  Google Scholar 

  • Heep M., Brandstätter B., Rieger U., Lehn N., Richter E., Rüsch-Gerdes S., Niemann S. (2001). Frequency of rpoB mutations inside and outside the cluster I region in rifampinresistant clinicalMycobacterium tuberculosis isolates. J. Clin. Microbiol., 39: 107–110.

    Article  CAS  PubMed  Google Scholar 

  • Heyman S.J., Brewer T.F., Wilson M.E., Fineberg H.V. (1999). The need for global action against multidrug-resistant tuberculosis. JAMA, 281: 2138–2140.

    Article  Google Scholar 

  • Huang H., Jin Q., Ma Y., Chen X., Zhang Y. (2002). Characterization of rpob mutations in rifampicin-resistantMycobacterium tuberculosis isolated in China. Tuberculosis, 82 (2–3): 79–83.

    Article  PubMed  Google Scholar 

  • Kapur V., Li L.L., Jordanescu S., Hamrick M.R., Wanger A., Kreiswirth B.N., Musser J.M. (1994). Characterization by automated DNA sequencing of mutations in the gene (rpoB) encoding the RNA polymerase β subunit in rifampin-resistantMycobacterium tuberculosis strains from New York City and Texas. J. Clin. Microbiol., 32: 1095–1098.

    CAS  PubMed  Google Scholar 

  • Lee A.S., Lim I.H., Tang L.L., Telenti A., Wong S.Y. (1999). Contribution of kasA analysis to detection of isoniazid resistantMycobacterium tuberculosis in Singapore. Antimicrob. Agents Ch., 43 (8): 2087–2089.

    CAS  Google Scholar 

  • Musser J.M. (1995). Antimicrobial agent resistance in mycobacteria: molecular genetic insights. Clin. Microbiol. Rev., 8: 496–514.

    CAS  PubMed  Google Scholar 

  • Nikolayevsky V., Brown T., Balabanova Y., Ruddy M., Fedorin I., Drobniewski F. (2004). Detection of mutations associated with isoniazid and rifampin resistance inMycobacterium tuberculosis isolates from Samara Region, Russian Federation. J. Clin. Microbiol., 42 (10): 4498–4502.

    Article  CAS  PubMed  Google Scholar 

  • Ouans S., Venter H., Dabbs E.R. (1997). Ribosylative inactivation of rifampicin byMycobacterium smegmatis is principal contributor to its low susceptibility to this antibiotic. Antimicrob. Agents Ch., 41: 2456–2460.

    Google Scholar 

  • Ozturk C.E., Sanic A., Kaya D., Ceyhan I. (2005). Molecular Analysis of isoniazid, rifampin and streptomycin resistance inMycobacterium tuberculosis isolates from patients with tuberculosis in Duzce, Turkey. Jpn. J. Infect. Dis., 58: 309–312.

    CAS  PubMed  Google Scholar 

  • Perrone C., Truchis P. (1995). Tuberculose multirésistante, epidémiologie, traitement, prévention et recherches diagnostiques. Méd. Interne Rev., 16: 547–552.

    Article  Google Scholar 

  • Pfyffer G.E. (2000). Drug-resistant tuberculosis: resistance mechanisms and rapid susceptibility testing. Schweiz. Med. Wochenschr., 130 (49): 1909–1913.

    CAS  PubMed  Google Scholar 

  • Ramaswammy S., Musser J.M. (1998). Molecular genetic basis of antimicrobial agent resistance inMycobacterium tuberculosis: update. Tubercle Lung Dis., 79: 3–29.

    Article  Google Scholar 

  • Ramaswammy S.V., Dou S.J., Rendon A., Yang Z., Cave M.D., Graviss E.A. (2004). Genotypic analysis of multidrug-resistantMycobacterium tuberculosis isolates from Monterry, Mexico. J. Med. Microbiol., 53: 107–113.

    Article  Google Scholar 

  • Ratsirahonana O., Rasolofo Razanamparany V., Rasolonavalona T., Rakotonirina V., Rakotoarisaonina A., Rakotoherisoa A., Ralamboson M., Cauchoix B., Rakotondramarina D., Ramarokoto H. (2002). Résistance deMycobacterium tuberculosis aux antituberculeux à Antananarivo en 2000. Institut Pasteur de Madagascar. Archives, 68 (1 and 2): 44–47.

    CAS  Google Scholar 

  • Rattan A., Kalia A., Ahmad N. (1998). Multidrug-resistantMycobacterium tuberculosis. Molecular perspectives. Emerg. Infect. Dis., 4 (2): 195–209.

    Article  CAS  PubMed  Google Scholar 

  • Ruiz M., Torres M.J., Llanos A.C., Arroyo A., Palomares J.C., Aznar J. (2004). Direct detection of rifampin-and isoniazid-resistantMycobacterium tuberculosis in auramine-rodamine-positive sputum specimens by Real-Time PCR. J. Clin. Microbiol., 42 (4): 1585–1589.

    Article  CAS  PubMed  Google Scholar 

  • Schilke K., Weyer K., Bretzel G., Amthor B., Brandt J., Sticht-Groh V., Fourie P.B., Haas W.H. (1999). Universal pattern ofrpoB gene mutations among multidrug-resistant isolates ofMycobacterium tuberculosis complex from Africa. Int. J. Tuber. Lung. Dis., 3: 620–626.

    CAS  Google Scholar 

  • Singh, P., Katoch V.M. (2006). Multi-drug resistant tuberculosis: current status and emerging tool for its management in India. J. Commun. Dis., 38 (3): 216–219.

    PubMed  Google Scholar 

  • Tanaka Y., Yazawa K., Dabbs E.R., Nishikawa K., Komaki H., Mikami Y., Miyaji M., Morisaki N., Iwasaki S. (1996). Different rifampicin inactivation mechanisms inNocardia and related taxa. Microbiol. Immunol., 40 (1): 1–4.

    CAS  PubMed  Google Scholar 

  • Telenti A., Imboden P., Marschesi F., Lowrie D., Cole S., Colston M.J., Matter L., Schopfer K., Bodmer T. (1993). Detection of rifampicin-rersistance mutations inMycobacterium tuberculosis. Lancet, 341: 647–650.

    Article  CAS  PubMed  Google Scholar 

  • Telenti A., Honoré N., Bernasconi C., March J., Ortega A., Heym B., Takiff H.E., Cole S.T. (1997). Genotypic assessment of isoniazid and rifampin resistance inMycobacterium tuberculosis: a blind study at reference laboratory level. J. Clin. Microbiol., 35 (3): 719–723.

    CAS  PubMed  Google Scholar 

  • Van Doorn H.R., Kuijper E.J., van der Ende A., Welten A.G., van Sooligen D., de Haas P.E., Dankert J. (2001). The susceptibility ofMycobacterium tuberculosis to isoniazid and the Arg→Leu mutation at codon 463 of katG are not associated. J. Clin. Microbiol., 39 (4):1591–1594.

    Article  PubMed  Google Scholar 

  • Van Rie A., Warren R., Mshanga I., Jordaan A.M., Vander Spuy G.D., Richardson M., Simpson J., Gie R.P., Enarson D.A., Beyer N., van Helden P.D., Victor T.C. (2001). Analysis for a limited number of gene codons can predict drug resistance ofMycobacterium tuberculosis in high-incidence community. J. Clin. Microbiol., 39: 636–641.

    Article  PubMed  Google Scholar 

  • Victor T.C., Jordaan A.M., van Rie A., van der Spuy G.D., Richardson M., van Helden P.D., Warren R. (1999). Detection of mutations in drug resistance genes ofMycobacterium tuberculosis by a dot-blot hybridisation strategy. Tubercle Lung. Dis., 79 (6): 343–348.

    Article  CAS  Google Scholar 

  • Victor T.C., van Helden P.D., Warren R. (2002). Prediction of drug resistance inMycobacterium tuberculosis: molecular mechanisms, tools, and applications. IUBMB Life, 53(4–5):231–7.

    Article  CAS  PubMed  Google Scholar 

  • WHO-World Health Organization (2002). Stop TB: Annual Report 2001. WHO/CDS/STB/2002.17, World Health Organization, Geneva, Switzerland.

    Google Scholar 

  • WHO-World Health Organization (2003). Global Tuberculosis Control 2003. World Health Organization, Geneva, Switzerland.

    Google Scholar 

  • WHO-World Health Organization (2007). Global Tuberculosis Control: Surveillance, Planning, Financing. WHO report 2007. WHO/HTM/TB/2007.376, World Health Organization, Geneva, Switzerland.

    Google Scholar 

  • Wilson T.M., Collins D.M. (1996). AhpC, a gene involved in izoniazid resistance of theMycobacterium tuberculosis complex. Mol. Microbiol., 19: 1025–1034.

    Article  CAS  PubMed  Google Scholar 

  • Wu X.Q., Lu Y., Zhang J.X., Liang J.Q., Li H.M., Zhang G.Y., Lü C.H., Ding B.C. (2006). Detection of the mutations in katG 315 and inhA-15 ofMycobacterium tuberculosis strains isolated from Chinese patients. Chin. Med. J. (Engl)., 119 (3): 230–233.

    CAS  Google Scholar 

  • Zhang Y., Heym B., Allen B., Young D., Cole S. (1992). The catalase-peroxidase gene and isoniazid resistance ofMycobacterium tuberculosis. Nature, 358: 591–593.

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Radia Sabouni.

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Sabouni, R., Kourout, M., Chaoui, I. et al. Molecular analysis of multidrug resistantMycobacterium tuberculosis isolates from Morocco. Ann. Microbiol. 58, 749–754 (2008). https://doi.org/10.1007/BF03175585

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