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Current Microbiology

, 53:329 | Cite as

Antibiotic Susceptibility of Helicobacter pylori Clinical Isolates: Comparative Evaluation of Disk-Diffusion and E-Test Methods

  • K. K. Mishra
  • S. Srivastava
  • A. Garg
  • A. AyyagariEmail author
Article

Abstract

Antimicrobial susceptibility of 25 Helicobacter pylori strains isolated from patients with acid peptic diseases were tested for in vitro sensitivity to commonly used antibiotics using disk-diffusion and E-test, methods. All strains tested were susceptible to tetracycline by E-test, with the minimum inhibitory concentration (MIC) values being <0.125 μg/ml for all strains except for 6 (<0.023 μg/ml). However 1 strain was resistant by disk-diffusion method. One strain was resistant to clarithromycin both by disk diffusion and E-test (MIC <48 μg/ml), and 1 strain was resistant only by disk diffusion. Only one strain was resistant to amoxicillin by disk diffusion and E-test (MIC >256 μg/ml). For ciprofloxacin, three strains were resistant by disk diffusion and two by E-test (MIC <32 μg/ml). Sixteen strains were resistant to metronidazole by disk diffusion and E-test (MIC ≥ 8 μg/ml), and 1 was resistant only by E-test (MIC <48 μg/ml). Overall, 64% of the strains were resistant to metronidazole. The MIC for metronidazole was also tested by agar-dilution method, and metronidazole resistant strains had an, MIC >8 μg/ml. The disk-diffusion method showed excellent correlation with E-test results; there was 100% agreement for amoxicillin a other antibiotics showed 90% to 95% accuracy. Disk diffusion is cheaper than E-test (approximately 2.6 cents vs. US$2.60), is easy to perform, and is a reliable method for testing H. pylori susceptibility to antimicrobial agents in the clinical microbiology laboratory.

Keywords

Minimum Inhibitory Concentration Metronidazole Amoxicillin Pylorus Infection Clarithromycin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

K.K.M. is thankful to S.R. Naik, Department of Gastroenterolgy, Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) (Lucknow, India) for extending clinical support to carry out the study.

Literature Cited

  1. 1.
    National Institute of Health Consensus conference (1994) Helicobacter pylori in peptic ulcer diseases. Development Panel on Helicobacter pylori in Peptic Ulcer Diseases. JAJMA 272:65–69CrossRefGoogle Scholar
  2. 2.
    Walsh JH, Peterson WL (1995) The treatment of Helicobacter pylori infection in the management of peptic ulcer diseases. N Eng J Med 333:984–991CrossRefGoogle Scholar
  3. 3.
    Debets-Ossenkopp YJ, Sparrius M, Kusters J (1996) Mechanism of clarithromycin resistance in clinical isolates of Helicobacter pylori. FEMS Microbiol Lett 142:37–42PubMedCrossRefGoogle Scholar
  4. 4.
    Megraud F, Occhialini A, Doennann HP (1997) Resistance of Helicobacter pylori to macrolides and nitroimidazole compounds—The current situation. J Physiol Pharmacol 48:25–38PubMedGoogle Scholar
  5. 5.
    Xia HX, Keane CT, O’Morain CA (1996) A 5 year survey of metronidazole and clarithromycine resistance in clinical isolates of Helicobacter pylori. Gut 39:(Suppl. 2) A6Google Scholar
  6. 6.
    Lopez-Brea M, Domingo D, Sanchez I Alarcon T (1997) Evolution of resistance to metronidazole and clarithromycin in Helicobacter pylori clinical isolates from Spain. J Antimicrob Chemother 40:279–281PubMedCrossRefGoogle Scholar
  7. 7.
    Van Zwet AA, De Boer WA, Schneeberger PM, Weel J, Thijs JC (1996) Prevalence of primary Helicobacter pylori resistance to metronidazole and clarithromycin in the Netherlands. Eur J Clin Microbiol Infect Dis 15:8614CrossRefGoogle Scholar
  8. 8.
    Van der Wouden EJ, van Zwet AA, Vosmear GDC Kleibeuker JR (1997) Rapid increase in the prevalence of metronidazole resistant Helicobacter pylori in the Netherlands. Emerg Infect Dis 3:385–389PubMedCrossRefGoogle Scholar
  9. 9.
    Midolo PD, Korman MG, Tumidge JD, Lambert JR (1996) Helicobacter pylori resistance to tetracycline. Lancet 347:1194–1195PubMedCrossRefGoogle Scholar
  10. 10.
    Megraud E (1997) Resistance of Helicobacter pylori to antibiotics. Aliment Pharmocol Ther 11:435–453CrossRefGoogle Scholar
  11. 11.
    Piccolomini R, Di Bonaventura G, Catamo G, Carbone F, Neri M (1997) Comparative evaluation of E-test, agar dilution and broth macrodilution for testing susceptibility of Helicobacter pylori strains to 20 antimicrobial agents. J Clin Microbiol 35:1842–1846PubMedGoogle Scholar
  12. 12.
    Mendonca S, Ecclissata C, Sartori MS, Godoy AP, Guerzoni RA, Dagger M, et al. (2000) Prevalence of Helicobacter pylori resistance to metronidazole clarithromycin, amoxicillin, tetracyclin and furazolidone in Brazil. Helicobacter 5:79–83PubMedCrossRefGoogle Scholar
  13. 13.
    Lahaie RG, Gaudreau C (2000) Helicobacter pylori antibiotic resistance: Trends over time. Can J Gastroenterol 14:895–899PubMedGoogle Scholar
  14. 14.
    Glupezynski Y, Labbe M, Hansen W, Yourassowsky E (1991) Evaluation of the E-test for quantitative antimicrobial susceptibility testing of Helicobacter pylori. L Clin Microbiol 29:2072–2075Google Scholar
  15. 15.
    Hachem CY, Clarridge JE, Reddy R, Graham DY (1996) Antimicrobial susceptibility testing of Helicobacter pylori. Comparison of E-test, broth microdilution, and disk diffusion for ampicillin, clarithromycin, and metronidazole. Diaga Microbiol Infect Dis 24:37–41CrossRefGoogle Scholar
  16. 16.
    Hirschi MA, Hirschi MM, Rotter ML (1993) Comparison of three methods for determination of the sensitivity of Helicobacter pylori to metronidazole. J Antimicrob Chemother 32:45–49Google Scholar
  17. 17.
    De Boer WA, Tytgat NJ (1996) How to treat Helicobacter pylori infection–should treatment strategies be based on testing bacterial susceptibility? A personal viewpoint. Eur J Gastroenterol Hepatol 8:709–716PubMedGoogle Scholar
  18. 18.
    Decross AJ, Maeshall BJ, McCallum RW, Hoffman SR, Barrett LJ, Guerrant RL (1993) Metronidazole susceptibility testing for Helicobacter pylori: Comparison of disc broth and agar dilution methods and their clinical relevance. J Clin Microbiol 31:1971–1974PubMedGoogle Scholar
  19. 19.
    Henriksen TH, Brorsom O, Schoyen R, Thoresen T (1996) Risk of lack of standardization of test to detect in vitro metronidazole resistance in Helicobacter pylori. Eur J Clin Microbiol Infect Dis 15:484–488PubMedCrossRefGoogle Scholar
  20. 20.
    Loo VG, Fallone CA, DeSouza E, Barkun AN (1996) In vitro susceptibility of Helicobacter pylori to ampicillin, clarithromycin, metronidazole, and omeprazole. J Antimicrob Chemother 40:881–883PubMedCrossRefGoogle Scholar
  21. 21.
    Chaves S, Gadanho M, Tenreiro R, Cabrita J (1993) Assessment of metronidazole susceptibility of Helicobacter pylori: statistical validation and error rate analysis of breakpoints determined by the disk diffusion test. Clin Microbiol 37:1628–1631PubMedGoogle Scholar
  22. 22.
    Cederbrant G, Kahlmeter G Ljunehh A (1993) E-test for antimicrobial susceptibility testing of Helicobacter pylori. J Antimicrob Chemother 31:65–71PubMedGoogle Scholar
  23. 23.
    Sharma S, Prasad KN, Chamoli D, Ayyagari A (1995) Antimicrobial susceptibility pattern biotyping of Helicobacter pylori isolates from patients with peptic ulcer diseases. Ind J Med Res 102:261–266Google Scholar
  24. 24.
    Mishra KK, Srivastava S, Dwivedi PP, Prasad KN, Ayyagari A (2002) ureCPCR of cagA gene based diagnosis of Helicobacter pylori infection and detection gastric biopsies. Ind J Pathol Microbiol 45:31–38Google Scholar
  25. 25.
    Mishra KK, Srivastava S, Dwivedi PP, Prasad KN, Ayyagari A (2002) Genotype of Helicobacter pylori isolated from various acid peptic diseases in and around Lucknow. Curr Sci 83:749–755Google Scholar
  26. 26.
    Bauer AW, Kirby WMM, Sherris JC, Truck M (1966) Antibiotic susceptibility testing by a single disc method. Am J Clin J Pathol 45:493Google Scholar
  27. 27.
    National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing. Ninth informational supplement M100–S10. Wayne, PA, National Committee for Clinical Laboratory Standards, 2000Google Scholar
  28. 28.
    Becx MC, Janssen AJ, Clasener HA, de Koning RW (1990) Metronidazole resistant Helicobacter pylori. Lancet 3:539–540CrossRefGoogle Scholar
  29. 29.
    Megraud F, Lehn N, Lind T, Bayerdroffer E, O’Morain C, Spiller R (1999) Antimicrobial therapy of Helicobacter pylori in a large multicenter trial: The MACH 2 study. Antimicrob Agents Chemother 43:2747–2752PubMedGoogle Scholar
  30. 30.
    McNulty CAM, Ent JC, Wise R (1998) Susceptibility of clinical isolates of Campylobacter pyloridis to 11 antimicrobial agents. Antimicrob Agents Chemother 28:837–838Google Scholar
  31. 31.
    Glupczynski Y, Burette A, DeKoster E (1990) Metronidazole resistance in Helicobacter pylori. Lancet 338:976–977CrossRefGoogle Scholar
  32. 32.
    Smith MA, Edwards DI (1995) The influence of microaerophilla and anaerobiosis on metronidazole uptake in Helicobacter pylori. J Antimicrob Chemother 36:453–461PubMedGoogle Scholar
  33. 33.
    Peterson WL, Graham DY, Marshall B, Blaser MJ, Genta RM, Klein PD (1993) Clarithromycin as monotherapy for eradication of Helicobacter pylori: A randomized, double-blind trail. Am J Gastroenterol 88:1860–1864PubMedGoogle Scholar
  34. 34.
    Megraud F (1998) Epidemiology and mechanism of antibiotic resistance in Helicobacter pylori. Gasroenterology 115:1278–1282CrossRefGoogle Scholar
  35. 35.
    Versalovic J, Shottridge D, Kibler K, Griffey MV, Beyer J, Flames RK (1996) Mutation in 23S rRNA are associated with clarithromycin resistance in Helicobacter pylori. Antimicrob Agents Chemother 40:377–380Google Scholar
  36. 36.
    Megraud F (1988) Antibiotic resistance in Helicobacter pylori infection. Br Med Bull 54:207–216Google Scholar
  37. 37.
    Megraud F (1997) Resistance of Helicobacter pylori to antibiotics. Aliment Pharmocol Ther 1:43–53CrossRefGoogle Scholar
  38. 38.
    Ani AE, Malu AO, Onah JA, Queiroz DM, Kirchner G, Rocha GA (1999) Antimicrobial susceptibility test of Helicobacter pylori isolated from Jos, Nigiria. Trans R Soc Trop Med Hyg 93:659–661PubMedCrossRefGoogle Scholar
  39. 39.
    Wolle K, Nilius M, Leodolter A, Muller WA, Molfertheiner P, Koning W (1998) Prevalence of Helicobacter pylori resistance to several antimicrobial agents in a region of Germany. Eur J Clin Microbiol Infect Dis 17: 519–521Google Scholar
  40. 40.
    Lopez BO, Moran VA, Ramirez AJA, Picazo de la GJJ (1998) Microbiologic diagnosis of Helicobacter pylori and its resistance to antibiotics. Rev Clin Esp 198:420–423Google Scholar
  41. 41.
    Bindayna KM (2001) Antibiotic susceptibilities of Helicobacter pylori. Saudi Med J 22:53–57PubMedGoogle Scholar
  42. 42.
    Casals JB, Pringler N (1991) Antifungal/antibacterial sensitivity testing using Neo-sensitabs. (9th Ed.) Taastrup, Denmark, Twisted roll diagnosesGoogle Scholar
  43. 43.
    Cederbrant G, Kahlmeter G, Liungh A (1993) E-test for antimicrobial susceptibility testing of Helicobacter pylori. J Antimicrob Chemother 31:65–71PubMedGoogle Scholar
  44. 44.
    Thyagarajan SP, Ray P, Das BK, Ayyagari A, Khan AA, Dharmalingam S, et al. (2003) Geographical differences in Antimicrobial resistance pattern of Helicobacter pylori clinical isolates from Indian patients: Multicentric study. J Gastroenterol Hepatol, 18:1373PubMedCrossRefGoogle Scholar
  45. 45.
    Kwon DH, Kirn JJ, Lee M, Yamaoka Y Kato M (2000) Isolation and characterization of tetracyclin resistant clinical isolates of Helicobacter pylori, Antimicroby Agents Chemother 44:3203–3205PubMedCrossRefGoogle Scholar
  46. 46.
    E-test technical guide 8. Susceptibility testing of Helicobacter pylori. AB Biodisk, Solna, SwedenGoogle Scholar
  47. 47.
    Dutta S, Chatopadhyay S, Patra R, De R, Ramamurthy T, Hembram J, et al. (2005) Most Helicobacter pylori strains in Kolkata in India are resistant to metronidazole but susceptible to other; commonly used for eradication and ulcer therapy. Ailment Pharmacol Ther 22:51–57CrossRefGoogle Scholar
  48. 48.
    Occhialini A, Urdaci M, Douect PF, Debear CM, Lamouliatte H, Megraud F (1997) Macrolid resistance in Helicobacter pylori: rapid detection and point mutations and assay of macrolid binding to ribosome. Antimicrob, Agents Chemother 41:2724–2728PubMedGoogle Scholar
  49. 49.
    Hachem CY, Claridge JE, Reddy R, Flamm R, Evans DG, Tanaka SK, Graham DY (1996) Antimicrobial susceptibility testing of Helicobacter pylori: Comparison of E-test, broth microdilution, and disk diffusion for ampicillin, clarithromycin, and metronidazole. Diagn Microbiol Infect Dis 24:37–41PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • K. K. Mishra
    • 1
  • S. Srivastava
    • 1
  • A. Garg
    • 1
  • A. Ayyagari
    • 1
    Email author
  1. 1.Department of MicrobiologySanjay Gandhi Postgraduate Institute of Medical SciencesLucknowIndia

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