Rifampin-resistance-associated mutations in the rifampin-resistance-determining region of the gene of clinical isolates in Shanghai, PR China Free

Abstract

Resistance to rifampin (RIF) in infection is associated with mutations in the gene coding for the β-subunit of RNA polymerase. The contribution of various mutations to the development and level of RIF resistance remains elusive.

Various mutations may be associated with differential levels of RIF resistance.

This study aimed to investigate the relationship between specific mutations and the MICs of RIF and rifabutin (RFB) against .

Of the 195 clinical isolates, 105 and 90 isolates were randomly selected from isolates resistant to RIF and sensitive to RIF, respectively. The MICs of 12 agents for isolates were determined using commercial Sensititre MIC plates and the broth microdilution method. Strains were screened for mutations by DNA extraction, gene amplification and DNA sequence analysis.

One hundred isolates (95.24 %) were found to have mutations in the RIF-resistance-determining region (RRDR) of the gene. Three mutations were identified in 90 RIF-susceptible isolates. Out of 105 isolates, 86 (81.90 %) were cross-resistant to both RIF and RFB. The most frequent mutation occurred at codons 450 and 445. We also found a novel nine-nucleotide (ATCATGCAT) deletion (between positions 1543 and 1551) in the gene in two strains (1.90 %) with resistance to RIF, but susceptibility to RFB. In addition, the mutation frequency at codon 450 was significantly higher in RIF-resistant/RFB-resistant (RIF/RFB) strains than in RIF/RFB strains (75.58 % versus 21.05 %, <0.01), whereas the mutation frequency at codon 435 was significantly lower in RIF/RFB strains than in RIF/RFB strains (1.16 % versus 26.32 %, <0.01).

Our data support previous findings, which reported that various mutations are associated with differential levels of RIF resistance. The specific mutations in the gene in RIF/RFB isolates differed from those in the RIF/RFB isolates. A novel deletion mutation in the RRDR might be associated with resistance to RIF, but not to RFB. Further clinical studies are required to investigate the efficacy of RFB in the treatment of infections caused by strains harbouring these mutations.

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2021-01-28
2024-03-29
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References

  1. WHO Global tuberculosis report.; 2018
  2. Steingart KR, Jotblad S, Robsky K, Deck D, Hopewell PC et al. Higher-dose rifampin for the treatment of pulmonary tuberculosis: a systematic review. Int J Tuberc Lung Dis 2011; 15:305–316[PubMed]
    [Google Scholar]
  3. Menzies D, Benedetti A, Paydar A, Martin I, Royce S et al. Effect of duration and intermittency of rifampin on tuberculosis treatment outcomes: a systematic review and meta-analysis. PLoS Med 2009; 6:e1000146 [View Article][PubMed]
    [Google Scholar]
  4. Gillespie SH. Evolution of drug resistance in Mycobacterium tuberculosis: clinical and molecular perspective. Antimicrob Agents Chemother 2002; 46:267–274 [View Article][PubMed]
    [Google Scholar]
  5. Musser JM. Antimicrobial agent resistance in mycobacteria: molecular genetic insights. Clin Microbiol Rev 1995; 8:496–514 [View Article][PubMed]
    [Google Scholar]
  6. Ohno H, Koga H, Kohno S, Tashiro T, Hara K. Relationship between rifampin MICs for and rpoB mutations of Mycobacterium tuberculosis strains isolated in Japan. Antimicrob Agents Chemother 1996; 40:1053–1056 [View Article][PubMed]
    [Google Scholar]
  7. Somoskovi A, Parsons LM, Salfinger M. The molecular basis of resistance to isoniazid, rifampin, and pyrazinamide in Mycobacterium tuberculosis . Respir Res 2001; 2:164–168 [View Article][PubMed]
    [Google Scholar]
  8. Bahrmand AR, Titov LP, Tasbiti AH, Yari S, Graviss EA. High-level rifampin resistance correlates with multiple mutations in the rpoB gene of pulmonary tuberculosis isolates from the Afghanistan border of Iran. J Clin Microbiol 2009; 47:2744–2750 [View Article][PubMed]
    [Google Scholar]
  9. Rigouts L, Gumusboga M, de Rijk WB, Nduwamahoro E, Uwizeye C et al. Rifampin resistance missed in automated liquid culture system for Mycobacterium tuberculosis isolates with specific rpoB mutations. J Clin Microbiol 2013; 51:2641–2645 [View Article][PubMed]
    [Google Scholar]
  10. Van Deun A, Barrera L, Bastian I, Fattorini L, Hoffmann H et al. Mycobacterium tuberculosis strains with highly discordant rifampin susceptibility test results. J Clin Microbiol 2009; 47:3501–3506 [View Article][PubMed]
    [Google Scholar]
  11. Van Deun A, Aung KJM, Bola V, Lebeke R, Hossain MA et al. Rifampin drug resistance tests for tuberculosis: challenging the gold standard. J Clin Microbiol 2013; 51:2633–2640 [View Article][PubMed]
    [Google Scholar]
  12. Marsili L, Pasqualucci CR, Vigevani A, Gioia B, Schioppacassi G et al. New rifamycins modified at positions 3 and 4. Synthesis, structure and biological evaluation. J Antibiot 1981; 34:1033–1038 [View Article][PubMed]
    [Google Scholar]
  13. Uzun M, Erturan Z, Anğ O. Investigation of cross-resistance between rifampin and rifabutin in Mycobacterium tuberculosis complex strains. Int J Tuberc Lung Dis 2002; 6:164–165[PubMed]
    [Google Scholar]
  14. Berrada ZL, Lin S-YG, Rodwell TC, Nguyen D, Schecter GF et al. Rifabutin and rifampin resistance levels and associated rpoB mutations in clinical isolates of Mycobacterium tuberculosis complex. Diagn Microbiol Infect Dis 2016; 85:177–181 [View Article][PubMed]
    [Google Scholar]
  15. Chikamatsu K, Mizuno K, Yamada H, Mitarai S. [Cross-resistance between rifampicin and rifabutin among multi-drug resistant Mycobacterium tuberculosis strains]. Kekkaku 2009; 84:631–633[PubMed]
    [Google Scholar]
  16. Clinical and Laboratory Standards Institute Performance Standards for Susceptibility Testing of Mycobacteria, Nocardia spp., and Other Aerobic Actinomycetes; approved astandard M62, 1st ed. CLSI; 2018
    [Google Scholar]
  17. Jamieson FB, Guthrie JL, Neemuchwala A, Lastovetska O, Melano RG et al. Profiling of rpoB mutations and MICs for rifampin and rifabutin in Mycobacterium tuberculosis . J Clin Microbiol 2014; 52:2157–2162 [View Article][PubMed]
    [Google Scholar]
  18. Abbadi SH, Sameaa GA, Morlock G, Cooksey RC. Molecular identification of mutations associated with anti-tuberculosis drug resistance among strains of Mycobacterium tuberculosis . Int J Infect Dis 2009; 13:673–678 [View Article][PubMed]
    [Google Scholar]
  19. Hillemann D, Weizenegger M, Kubica T, Richter E, Niemann S. Use of the genotype MTBDR assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis complex isolates. J Clin Microbiol 2005; 43:3699–3703 [View Article][PubMed]
    [Google Scholar]
  20. Islam MM, Hameed HMA, Mugweru J, Chhotaray C, Wang C et al. Drug resistance mechanisms and novel drug targets for tuberculosis therapy. J Genet Genomics 2017; 44:21–37 [View Article][PubMed]
    [Google Scholar]
  21. Moghazeh SL, Pan X, Arain T, Stover CK, Musser JM et al. Comparative antimycobacterial activities of rifampin, rifapentine, and KRM-1648 against a collection of rifampin-resistant Mycobacterium tuberculosis isolates with known rpoB mutations. Antimicrob Agents Chemother 1996; 40:2655–2657 [View Article][PubMed]
    [Google Scholar]
  22. Tan Y, Hu Z, Zhao Y, Cai X, Luo C et al. The beginning of the rpoB gene in addition to the rifampin resistance determination region might be needed for identifying rifampin/rifabutin cross-resistance in multidrug-resistant Mycobacterium tuberculosis isolates from southern China. J Clin Microbiol 2012; 50:81–85 [View Article][PubMed]
    [Google Scholar]
  23. Torres MJ, Criado A, Gónzalez N, Palomares JC, Aznar J. Rifampin and isoniazid resistance associated mutations in Mycobacterium tuberculosis clinical isolates in Seville, Spain. Int J Tuberc Lung Dis 2002; 6:160–163[PubMed]
    [Google Scholar]
  24. Chen J, Peng P, Du Y, Ren Y, Chen L et al. Early detection of multidrug- and pre-extensively drug-resistant tuberculosis from smear-positive sputum by direct sequencing. BMC Infect Dis 2017; 17:300 [View Article][PubMed]
    [Google Scholar]
  25. Titov LP, Zakerbostanabad S, Slizen V, Surkova L, Taghikhani M et al. Molecular characterization of rpoB gene mutations in rifampicine-resistant Mycobacterium tuberculosis isolates from tuberculosis patients in Belarus. Biotechnol J 2006; 1:1447–1452 [View Article][PubMed]
    [Google Scholar]
  26. Yu X-li, Wen Z-lu, Chen G-zhan, Li R, Ding B-bing et al. Molecular characterization of multidrug-resistant Mycobacterium tuberculosis isolated from south-central in China. J Antibiot 2014; 67:291–297 [View Article][PubMed]
    [Google Scholar]
  27. Zaczek A, Brzostek A, Augustynowicz-Kopec E, Zwolska Z, Dziadek J. Genetic evaluation of relationship between mutations in rpoB and resistance of Mycobacterium tuberculosis to rifampin. BMC Microbiol 2009; 9:10 [View Article][PubMed]
    [Google Scholar]
  28. Makhado NA, Matabane E, Faccin M, Pinçon C, Jouet A et al. Outbreak of multidrug-resistant tuberculosis in South Africa undetected by WHO-endorsed commercial tests: an observational study. Lancet Infect Dis 2018; 18:1350–1359 [View Article][PubMed]
    [Google Scholar]
  29. Siu GKH, Zhang Y, Lau TCK, Lau RWT, Ho P-L et al. Mutations outside the rifampicin resistance-determining region associated with rifampicin resistance in Mycobacterium tuberculosis . J Antimicrob Chemother 2011; 66:730–733 [View Article][PubMed]
    [Google Scholar]
  30. Sanchez-Padilla E, Merker M, Beckert P, Jochims F, Dlamini T et al. Detection of drug-resistant tuberculosis by Xpert MTB/RIF in Swaziland. N Engl J Med 2015; 372:1181–1182 [View Article][PubMed]
    [Google Scholar]
  31. Donnabella V, Martiniuk F, Kinney D, Bacerdo M, Bonk S et al. Isolation of the gene for the beta subunit of RNA polymerase from rifampicin-resistant Mycobacterium tuberculosis and identification of new mutations. Am J Respir Cell Mol Biol 1994; 11:639–643 [View Article][PubMed]
    [Google Scholar]
  32. Huang H, Jin Q, Ma Y, Chen X, Zhuang Y. Characterization of rpoB mutations in rifampicin-resistant Mycobacterium tuberculosis isolated in China. Tuberculosis 2002; 82:79–83 [View Article][PubMed]
    [Google Scholar]
  33. Williams DL, Spring L, Collins L, Miller LP, Heifets LB et al. Contribution of rpoB mutations to development of rifamycin cross-resistance in Mycobacterium tuberculosis . Antimicrob Agents Chemother 1998; 42:1853–1857 [View Article][PubMed]
    [Google Scholar]
  34. Kapur V, Li LL, Iordanescu S, Hamrick MR, Wanger A et al. Characterization by automated DNA sequencing of mutations in the gene (rpoB) encoding the RNA polymerase beta subunit in rifampin-resistant Mycobacterium tuberculosis strains from New York City and Texas. J Clin Microbiol 1994; 32:1095–1098 [View Article][PubMed]
    [Google Scholar]
  35. Bodmer T, Zürcher G, Imboden P, Telenti A. Mutation position and type of substitution in the beta-subunit of the RNA polymerase influence in-vitro activity of rifamycins in rifampicin-resistant Mycobacterium tuberculosis . J Antimicrob Chemother 1995; 35:345–348 [View Article][PubMed]
    [Google Scholar]
  36. Herrera L, Jiménez S, Valverde A, García-Aranda MA, Sáez-Nieto JA. Molecular analysis of rifampicin-resistant Mycobacterium tuberculosis isolated in Spain (1996-2001). description of new mutations in the rpoB gene and review of the literature. Int J Antimicrob Agents 2003; 21:403–408 [View Article][PubMed]
    [Google Scholar]
  37. Hirano K, Abe C, Takahashi M. Mutations in the rpoB gene of rifampin-resistant Mycobacterium tuberculosis strains isolated mostly in Asian countries and their rapid detection by line probe assay. J Clin Microbiol 1999; 37:2663–2666 [View Article][PubMed]
    [Google Scholar]
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