Susceptibility Testing of Helicobacter pylori to Metronidazole and DNA Fingerprints of Resistant Strains in Singapore

  • HUA, JIESONG (NUS H. pylori Research Group, Departments of Microbiology National University of Singapore) ;
  • KHAY-GUAN YEOH (NUS H. pylori Research Group, Department of Medicine, National University of Singapore) ;
  • PENGYUAN ZHENG (NUS H. Pylori Research Group, Departments of Microbiology National University of Singapore) ;
  • HAN CHONG NG (NUS H. Pylori Research Group, Departments of Microbiology National University of Singapore) ;
  • BOW HO (NUS H. Pylori Research Group, Departments of Microbiology National University of Singapore)
  • Published : 1999.06.01

Abstract

Susceptibility of 61 strains of Helicobacter pylori to metronidazole was examined by both the disk diffusion method using a cut-off of 15㎜ for resistance and the E test with a cut-off of 8㎎/l. The MIC/sub 50/ and MIC/sub 90/ by the E test were 2 ㎎/l and 256㎎/l, respectively. Metronidazole resistance was found in 22 (36%) out of the 61 H. pylori strains by the E test and in three additional strains by the disk diffusion method. Amongst the latter three isolates, the MICs by the E test were 4 ㎎/l, 6㎎/l, and 6㎎/l, respectively. These figures are one log₂ or half log₂ dilution lower than the cut-off of 8㎎/l recommended as resistance for the E test. All 22 metronidazole resistant H. pylori isolates by the E test that were subjected to random amplified polymorphic DNA (RAPD) fingerprinting showed different DNA fingerprints. Interestingly, >90% of resistant isolates possess two common DNA bands of 0.4 and 0.9 kb. This study demonstrates that the results of the disk diffusion method for testing H. pylori susceptibility to metronidazole correlates well with that of the E test. The criteria for interpretation need to be internationally standardized so that the results from different centers can be compared.

Keywords

References

  1. Nucl. Acids. Res. v.20 DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting Akopyanz, N.;N. O. Bukanov;T. U. Westblom;S. Kresovich;D. E. Berg
  2. J. Clin. Microbiol. v.36 Discrepancies between E-test and agar dilution methods for testing metronidazole susceptibility of Helicobacter pylori Alarcon, T.;D. Domingo;M. Lopez-Brea
  3. Biochem. Biophys. Res. Commun. v.236 Isolation of a genetic locus associated with metronidazole resistance in Helicobacter pylori Chang, K. C.;S. W. Ho;J. C. Yang;J. T. Wang
  4. Gut v.38 Prevalence of metronidazole resistant Helicobacter pylori strains among Chinese peptic ulcer disease patients and normal controls in Hong Kong Ching, C. K.;K. P. Leung;R. W. Yung;S. K. Lam;B. C. Wong;K. C. Lai;C. L. Lai
  5. J. Clin. Microbiol. v.31 Metronidazole susceptibility testing for Helicobacter pylori : Comprison of disk, broth, and agar dilution methods and their clinical relevance DeCross, A. J.;B. J. Marshall;R. W. McCallum;S. R. Hoffman;L. J. Barrett;R. L. Guerrant
  6. British Med. J. v.316 Varying efficacy of Helicobacter pylori eradication regimens: cost effectiveness study using a decision analysis model Duggan, A. E.;K. Tolley;C. J. Hawkey;R. F. Logan
  7. J. Antimicrob, Chemother. v.31 Nitroimidazole drugs-action and resistance mechanisms. II. Mechanisms of resistance Edwards, D. I.
  8. Mol. Microbiol. v.28 Metronidazole resistance in Helicobacter pylori is due to null mutations in a gene (rdxA) that encodes an oxygen-insensitive NADPH nitroeductase Goodwin, A.;D. Kersulyte;G. Sisson;S. J. Veldhuyzen van Zanten;D. E. Berg;P. S. Hoffman
  9. Diagn. Microbiol. Infect. Dis v.24 Antimicrobial susceptibility testing of Helicobacter pylori.Comparison of E-test, broth microdilution, and disk diffusion for ampicillin, clarithromycin, and metronidazole Hachem, C. Y.;J. E. Clarridge;R. Reddy;R. Flamm;D. G. Evans;S. K. Tanaka;D. Y. Graham
  10. Helicobacter pylori Technique for Clinical Diagnosis and Basic Research PCR-based RAPD (random amplified polymorphic DNA) fingerprinting of clinical isolates of Helicobacter pylori Hua, J.;C. Birac;F. Megraud;A. Lee(ed.);F. Megraud(ed.)
  11. Microbios v.87 Is the coccoid form of Helicobacter pylori viable? Hua, J.;B. Ho
  12. World J. Gastroenterol v.4 Helicobacter pylori acquisition of metronidazole resistance by natural transformation in vitro Hua, J.;P. Y. Zheng;K. F. Teo;M. M. Khin;B. Ho
  13. Gut v.41 Does treatment of Helicobacter pylori with antibiotics alone heal duodenal ulcer? A randomised double blind placebo controlled study Lam, S. K.;C. K. Ching;K. C. Lai;B. C. Wong;C. L. Lai;C. K. Chan;L. Ong
  14. J. Gastroenterol. Hepatol. v.13 Report of the 1997 Asia Pacific Consensus Conference on the management of Helicobacter pylori infection Lam, S. K.;N. J. Talley
  15. Helicobacter v.1 An increase in Helicobacter pylori strains resistant to metronidazole: A five-year study Ling, T, K.;A. F. Cheng;J. J. Sung;P. Y. Yiu;S. S. Chung
  16. J. Infect. Dis. v.161 Natural competence for genetic transformation in Campylobacter pylori Nedenskov-Sorensen, P.;G. Bukholm;K. Bovre
  17. Scand. J. Infect. Dis. v.26 Impact of metronidazole resistance on the eradication of Helicobacter pylori Noach, L. A.;W. L. Langenberg;M. A. Bertola;J. Dankert;G. N. Tytgat
  18. J. Med. Microbiol. v.38 Biotype and molecular fingerprints of metronidazole-resistant strains of Helicobacter pylori from antral gastric mucosa Owen, R. J.;G. D. Bell;M. Desai;M. Moreno;P. W. Gant;P. H. Jones;D. Linton
  19. J. Clin. Microbiol. v.35 Comparative evaluation of the E test, agar dilution , and broth microdilution for testing susceptibilities of Helicobacter pylori strains to 20 antimicrobial agents Piccolomini, R.;G. Di Bonaventura;G. Catamo;F. Carbone;M. Neri
  20. Helicobacter v.2 Omeprazole-based dual and triple therapy for the treatment of Helicobacter pylori infection in peptic ulcer disease : A randomized trial Pieramico, O.;M. V. Zanetti;M. Innerhofer;P. Malfertheiner
  21. Antimicrob. Agents Chemother v.36 Role of metronidazole resistance in therapy of Helicobacter pylori infections Rautelin, H.;K. Seppala;O. V. Renkonen;U. Vainio;T. U. Kosunen
  22. J. Antimicrob. Chemother. v.3 The susceptibillity of Helicobacter pylori to 12 antimicrobial agents, omeprazole and bismuth salts Rubinstein, G.;K. Dunkin;A. J. Howard
  23. Am. J. Gastroenterol. v.93 Triple versus dual therapy for eradicating Helicobacter pylori and preventing ulcer recurrence: A randomized, double-blind, multicenter study fo lansoprazole, clarithromycin, and/or amoxicillin in different dosing regimens Schwartz, H.;R. Krause;B. Sahba;M. Haber;A. Weissfeld;P. Rose;N. Siepman;J. Freston
  24. Nature v.388 The complete genome sequance of the gastic pathogen: Helicobacter pylori Tomb, J. F.;O. White;A. R. Kerlavage;R. A. Clayton;G. G. Sutton;R. D. Fleischmann;K. A. Ketchum;H. P. Klenk;S. Gill;B. A. Dougherty;K. Nelson;J. Quackenbush;L. Zhou;E. F. Kirkness;S. Peterson;B. Loftus;D. Richardson;R. Dodson;H. G. Khalak; A. Glodek;K. McKenney;L. M. Fitzegerald;N. Lee;M. D. Adams;J. C. Venter;D. E. Berg;J. D. Gocayne;T. R. Utterback;J. D. Peterson;J. M. Kelly;M. D. Cotton;J. M. Weldman;C. Fujii;C. Bowman; L. Watthey;E. Wallin;W. S. Hayes;M. Borodovsky;P. D. Karp;H. O. Smith;C. M. Fraser;J. C. Venter
  25. J. Gen. Microbiol. v.139 Transformation of Helicobacter pylori by chromosomal mentronidazole resistance and by a plasmid with a selectable chloramphenicol resistance marker Wang, Y.;K. P. Roos;D. E. Taylor
  26. Antimicrob. Agents Chemother. v.38 Standardization of disk diffusion test and its clinicla significance for susceptiblity testing of metronidazole against Helicobacter pylori Xia, H.;C. T. Keane;S. Beattie;C. A. O'Morain