1887

Journal of Medical Microbiology logo

Volume 53, Issue 2

Frameshift mutations in occur frequently and do not provide a reliable marker for metronidazole resistance in UK isolates of Free

Abstract

Mutations in the NAD(P)H flavin oxidoreductase gene () are thought to contribute to the development of metronidazole resistance in . To test this further, 44 sequences in 18 patient isolate sets of were examined including a unique collection comprising separated Mtz-sensitive (Mtz) and Mtz-resistant (Mtz) subpopulations pre-treatment and matched Mtz strains post-treatment. Sequences of contained frameshift mutations that led to premature protein truncation in at least one strain from most (17/18) patient sets. These mutations were present in all strains, irrespective of Mtz resistotype in 13/18 patients. Frameshift due to a single adenine deletion at nucleotide 53 was the most common mutation and was present in isolates from 11/18 patients. A novel real-time (LightCycler) PCR-based probe hybridization melting-point assay applied to a further 119 isolates confirmed that the frameshift-53 mutation occurred frequently, in 20 % of isolates, and could be present in Mtz as well as Mtz strains (42 % vs 58 %). This study demonstrates that frameshift mutations occur in Mtz strains as well as in Mtz strains, and are thus unlikely to cause Mtz resistance.

  • Received:
  • Accepted:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.05342-0
2004-02-01
2024-04-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/53/2/JM530210.html?itemId=/content/journal/jmm/10.1099/jmm.0.05342-0&mimeType=html&fmt=ahah

References

  1. Alarcon T., Domingo D., Lopez-Brea M. 1999; Antibiotic resistance problems with Helicobacter pylori . Int J Antimicrob Agents 12:19–26 [CrossRef]
     [Google Scholar]
  2. Anonymous 1994; NIH Consensus Conference. Helicobacter pylori in peptic ulcer disease. NIH Consensus Development Panel on Helicobacter pylori in Peptic Ulcer Disease. JAMA (J Am Med Assoc) 272:65–69 [CrossRef]
     [Google Scholar]
  3. Appelmelk B. J., Martin S. L., Monteiro M. A. & 10 other authors; 1999; Phase variation in Helicobacter pylori lipopolysaccharide due to changes in the lengths of poly(C) tracts in alpha3-fucosyltransferase genes. Infect Immun 67:5361–5366
     [Google Scholar]
  4. Chisholm S. A., Owen R. J. 2003; Mutations in Helicobacter pylori rdxA gene sequences may not contribute to metronidazole resistance. J Antimicrob Chemother 51:995–999 [CrossRef]
     [Google Scholar]
  5. Chisholm S. A., Owen R. J., Teare E. L., Saverymuttu S. 2001; PCR-based diagnosis of Helicobacter pylori infection and real-time determination of clarithromycin resistance directly from human gastric biopsy samples. J Clin Microbiol 39:1217–1220 [CrossRef]
     [Google Scholar]
  6. Dore M. P., Leandro G., Realdi G., Sepulveda A. R., Graham D. Y. 2000; Effect of pretreatment antibiotic resistance to metronidazole and clarithromycin on outcome of Helicobacter pylori therapy: a meta-analytical approach. Dig Dis Sci 45:68–76 [CrossRef]
     [Google Scholar]
  7. Goodwin A., Kersulyte D., Sisson G., Veldhuyzen van Zanten S. J., Berg D. E., Hoffman P. S. 1998; Metronidazole resistance in Helicobacter pylori is due to null mutations in a gene ( rdxA ) that encodes an oxygen-insensitive NADPH nitroreductase. Mol Microbiol 28:383–393 [CrossRef]
     [Google Scholar]
  8. Helicobacter and Cancer Collaborative Group 2001; Gastric cancer and Helicobacter pylori : a combined analysis of 12 case control studies nested within prospective cohorts. Gut 49:347–353 [CrossRef]
     [Google Scholar]
  9. Jenks P. J., Edwards D. I. 2002; Metronidazole resistance in Helicobacter pylori . Int J Antimicrob Agents 19:1–7 [CrossRef]
     [Google Scholar]
  10. Jenks P. J., Ferrero R. L., Labigne A. 1999; The role of the rdxA gene in the evolution of metronidazole resistance in Helicobacter pylori . J Antimicrob Chemother 43:753–758 [CrossRef]
     [Google Scholar]
  11. Jeong J. Y., Mukhopadhyay A. K., Dailidiene D. & 20 other authors; 2000; Sequential inactivation of rdxA (HP0954) and frxA (HP0642) nitroreductase genes causes moderate and high-level metronidazole resistance in Helicobacter pylori . J Bacteriol 182:5082–5090 [CrossRef]
     [Google Scholar]
  12. Jeong J. Y., Mukhopadhyay A. K., Akada J. K., Dailidiene D., Hoffman P. S., Berg D. E. 2001; Roles of FrxA and RdxA nitroreductases of Helicobacter pylori in susceptibility and resistance to metronidazole. J Bacteriol 183:5155–5162 [CrossRef]
     [Google Scholar]
  13. Josenhans C., Eaton K. A., Thevenot T., Suerbaum S. 2000; Switching of flagellar motility in Helicobacter pylori by reversible length variation of a short homopolymeric sequence repeat in fliP , a gene encoding a basal body protein. Infect Immun 68:4598–4603 [CrossRef]
     [Google Scholar]
  14. Kwon D. H., El-Zaatari F. A., Kato M., Osato M. S., Reddy R., Yamaoka Y., Graham D. Y. 2000a; Analysis of rdxA and involvement of additional genes encoding NAD(P)H flavin oxidoreductase (FrxA) and ferredoxin-like protein (FdxB) in metronidazole resistance of Helicobacter pylori . Antimicrob Agents Chemother 44:2133–2142 [CrossRef]
     [Google Scholar]
  15. Kwon D. H., Kato M., El-Zaatari F. A., Osato M. S., Graham D. Y. 2000b; Frame-shift mutations in NAD(P)H flavin oxidoreductase encoding gene ( frxA ) from metronidazole resistant Helicobacter pylori ATCC43504 and its involvement in metronidazole resistance. FEMS Microbiol Lett 188:197–202 [CrossRef]
     [Google Scholar]
  16. Kwon D. H., Hulten K., Kato M., Kim J. J., Lee M., El-Zaatari F. A., Osato M. S., Graham D. Y. 2001a; DNA sequence analysis of rdxA and frxA from 12 pairs of metronidazole-sensitive and -resistant clinical Helicobacter pylori isolates. Antimicrob Agents Chemother 45:2609–2615 [CrossRef]
     [Google Scholar]
  17. Kwon D. H., Lee M., Kim J. J., Kim J. G., El-Zaatari F. A., Osato M. S., Graham D. Y. 2001b; Furazolidone- and nitrofurantoin-resistant Helicobacter pylori : prevalence and role of genes involved in metronidazole resistance. Antimicrob Agents Chemother 45:306–308 [CrossRef]
     [Google Scholar]
  18. Maeda S., Yoshida H., Matsunaga H., Ogura K., Kawamata O., Shiratori Y., Omata M. 2000; Detection of clarithromycin-resistant Helicobacter pylori strains by a preferential homoduplex formation assay. J Clin Microbiol 38:210–214
     [Google Scholar]
  19. Marais A., Bilardi C., Cantet F., Mendz G. L., Megraud F. 2003; Characterization of the genes rdxA and frxA involved in metronidazole resistance in Helicobacter pylori . Res Microbiol 154:137–144 [CrossRef]
     [Google Scholar]
  20. Matsumura M., Hikiba Y., Ogura K., Togo G., Tsukuda I., Ushikawa K., Shiratori Y., Omata M. 2001; Rapid detection of mutations in the 23S rRNA gene of Helicobacter pylori that confers resistance to clarithromycin treatment to the bacterium. J Clin Microbiol 39:691–695 [CrossRef]
     [Google Scholar]
  21. Megraud F., Lehn N., Lind T. & 7 other authors; 1999; Antimicrobial susceptibility testing of Helicobacter pylori in a large multicenter trial: the MACH 2 study. Antimicrob Agents Chemother 43:2747–2752
     [Google Scholar]
  22. Mendz G. L., Megraud F. 2002; Is the molecular basis of metronidazole resistance in microaerophilic organisms understood?. Trends Microbiol 10:370–375 [CrossRef]
     [Google Scholar]
  23. Mukhopadhyay A. K., Jeong J. Y., Dailidiene D., Hoffman P. S., Berg D. E. 2003; The fdxA ferredoxin gene can down-regulate frxA nitroreductase gene expression and is essential in many strains of Helicobacter pylori . J Bacteriol 185:2927–2935 [CrossRef]
     [Google Scholar]
  24. Owen R. J., Xerry J. 2003; Tracing clonality of Helicobacter pylori infecting family members from analysis of DNA sequences of three housekeeping genes ( ureI , atpA and ahpC ), deduced amino acid sequences, and pathogenicity-associated markers ( cagA and vacA ). J Med Microbiol 52:515–524 [CrossRef]
     [Google Scholar]
  25. Peck B., Ortkamp M., Diehl K. D., Hundt E., Knapp B. 1999; Conservation, localization and expression of HopZ, a protein involved in adhesion of Helicobacter pylori . Nucleic Acids Res 27:3325–3333 [CrossRef]
     [Google Scholar]
  26. Sisson G., Goodwin A., Raudonikiene A., Hughes N. J., Mukhopadhyay A. K., Berg D. E., Hoffman P. S. 2002; Enzymes associated with reductive activation and action of nitazoxanide, nitrofurans, and metronidazole in Helicobacter pylori . Antimicrob Agents Chemother 46:2116–2123 [CrossRef]
     [Google Scholar]
  27. Tankovic J., Lamarque D., Delchier J. C., Soussy C. J., Labigne A., Jenks P. J. 2000; Frequent association between alteration of the rdxA gene and metronidazole resistance in French and North African isolates of Helicobacter pylori . Antimicrob Agents Chemother 44:608–613 [CrossRef]
     [Google Scholar]
  28. Trebesius K., Panthel K., Strobel S., Vogt K., Faller G., Kirchner T., Kist M., Heesemann J., Haas R. 2000; Rapid and specific detection of Helicobacter pylori macrolide resistance in gastric tissue by fluorescent in situ hybridisation. Gut 46:608–614 [CrossRef]
     [Google Scholar]
  29. van der Wouden E. J., Thijs J. C., van Zwet A. A., Sluiter W. J., Kleibeuker J. H. 1999; The influence of in vitro nitroimidazole resistance on the efficacy of nitroimidazole-containing anti- Helicobacter pylori regimens: a meta-analysis. Am J Gastroenterol 94:1751–1759 [CrossRef]
     [Google Scholar]
  30. Wang G., Wilson T. J., Jiang Q., Taylor D. E. 2001; Spontaneous mutations that confer antibiotic resistance in Helicobacter pylori . Antimicrob Agents Chemother 45:727–733 [CrossRef]
     [Google Scholar]
  31. Wilson K. 1987; Preparation of genomic DNA from bacteria. In Current Protocols in Molecular Biology pp 241–242 Edited by Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Smith J. A., Seidman J. G., Struhl K. New York: Wiley;
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.05342-0
Loading
Frameshift mutations in frxA occur frequently and do not provide a reliable marker for metronidazole resistance in UK isolates of Helicobacter pylori
J. Med. Microbiol. 53, 135 (2004); https://doi.org/10.1099/jmm.0.05342-0
/content/journal/jmm/10.1099/jmm.0.05342-0
/content/journal/jmm/10.1099/jmm.0.05342-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error