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. 1994 May;32(5):1203–1210. doi: 10.1128/jcm.32.5.1203-1210.1994

Comparison of PCR-based restriction length polymorphism analysis of urease genes with rRNA gene profiling for monitoring Helicobacter pylori infections in patients on triple therapy.

R J Owen 1, J Bickley 1, A Hurtado 1, A Fraser 1, R E Pounder 1
PMCID: PMC263645  PMID: 7914204

Abstract

Multiple isolates of Helicobacter pylori from antral biopsies of nine patients were examined by DNA fingerprinting. Analysis of rRNA gene patterns and HaeIII restriction fragment length polymorphism of PCR-amplified urease genes were compared and used to study colonization before and after failed triple therapy. H. pylori isolates from a single biopsy shared the same HaeIII DNA fingerprint regardless of the isolation method (plate or broth). DNA pattern types of paired strains of H. pylori were distinct between patients and were not grossly affected by treatment except for one patient with an altered strain type. H. pylori infections were generally associated with several subpopulations of strains, evident from the subtypic variation before and after treatment, detectable by both DNA fingerprinting methods. The urease gene patterns also provided evidence that some cultures of H. pylori probably contained a mixture of genomic subtypes. The study suggests that triple therapy has the effect either of inducing minor genomic variations or of changing the proportions of different subtypes of H. pylori. It was concluded that urease gene profiling provides a simple yet reliable method of establishing whether treatment failures are attributable to incomplete eradication of H. pylori.

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Selected References

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  1. Akopyanz N., Bukanov N. O., Westblom T. U., Berg D. E. PCR-based RFLP analysis of DNA sequence diversity in the gastric pathogen Helicobacter pylori. Nucleic Acids Res. 1992 Dec 11;20(23):6221–6225. doi: 10.1093/nar/20.23.6221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beji A., Vincent P., Darchis I., Husson M. O., Cortot A., Leclerc H. Evidence of gastritis with several Helicobacter pylori strains. Lancet. 1989 Dec 9;2(8676):1402–1403. doi: 10.1016/s0140-6736(89)92020-5. [DOI] [PubMed] [Google Scholar]
  3. Bell G. D., Powell K. U., Burridge S. M., Spencer G., Bolton G., Purser K., Brooks S., Prosser S., Harrison G., Gant P. W. Short report: omeprazole plus antibiotic combinations for the eradication of metronidazole-resistant Helicobacter pylori. Aliment Pharmacol Ther. 1992 Dec;6(6):751–758. doi: 10.1111/j.1365-2036.1992.tb00740.x. [DOI] [PubMed] [Google Scholar]
  4. Bell G. D., Powell K., Burridge S. M., Pallecaros A., Jones P. H., Gant P. W., Harrison G., Trowell J. E. Experience with 'triple' anti-Helicobacter pylori eradication therapy: side effects and the importance of testing the pre-treatment bacterial isolate for metronidazole resistance. Aliment Pharmacol Ther. 1992 Aug;6(4):427–435. doi: 10.1111/j.1365-2036.1992.tb00556.x. [DOI] [PubMed] [Google Scholar]
  5. Clayton C. L., Kleanthous H., Morgan D. D., Puckey L., Tabaqchali S. Rapid fingerprinting of Helicobacter pylori by polymerase chain reaction and restriction fragment length polymorphism analysis. J Clin Microbiol. 1993 Jun;31(6):1420–1425. doi: 10.1128/jcm.31.6.1420-1425.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Costas M. Numerical analysis of sodium dodecyl sulphate-polyacrylamide gel electrophoretic protein patterns for the classification, identification and typing of medically important bacteria. Electrophoresis. 1990 May;11(5):382–391. doi: 10.1002/elps.1150110507. [DOI] [PubMed] [Google Scholar]
  7. Foxall P. A., Hu L. T., Mobley H. L. Use of polymerase chain reaction-amplified Helicobacter pylori urease structural genes for differentiation of isolates. J Clin Microbiol. 1992 Mar;30(3):739–741. doi: 10.1128/jcm.30.3.739-741.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fraser A. G., Bickley J., Owen R. J., Pounder R. E. DNA fingerprints of Helicobacter pylori before and after treatment with omeprazole. J Clin Pathol. 1992 Dec;45(12):1062–1065. doi: 10.1136/jcp.45.12.1062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Graham D. Y. Campylobacter pylori and peptic ulcer disease. Gastroenterology. 1989 Feb;96(2 Pt 2 Suppl):615–625. doi: 10.1016/s0016-5085(89)80057-5. [DOI] [PubMed] [Google Scholar]
  10. Graham D. Y., Lew G. M., Klein P. D., Evans D. G., Evans D. J., Jr, Saeed Z. A., Malaty H. M. Effect of treatment of Helicobacter pylori infection on the long-term recurrence of gastric or duodenal ulcer. A randomized, controlled study. Ann Intern Med. 1992 May 1;116(9):705–708. doi: 10.7326/0003-4819-116-9-705. [DOI] [PubMed] [Google Scholar]
  11. Labigne A., Cussac V., Courcoux P. Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol. 1991 Mar;173(6):1920–1931. doi: 10.1128/jb.173.6.1920-1931.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Langenberg W., Rauws E. A., Widjojokusumo A., Tytgat G. N., Zanen H. C. Identification of Campylobacter pyloridis isolates by restriction endonuclease DNA analysis. J Clin Microbiol. 1986 Sep;24(3):414–417. doi: 10.1128/jcm.24.3.414-417.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lopez C. R., Owen R. J., Desai M. Differentiation between isolates of Helicobacter pylori by PCR-RFLP analysis of urease A and B genes and comparison with ribosomal RNA gene patterns. FEMS Microbiol Lett. 1993 Jun 1;110(1):37–43. doi: 10.1111/j.1574-6968.1993.tb06292.x. [DOI] [PubMed] [Google Scholar]
  14. Majewski S. I., Goodwin C. S. Restriction endonuclease analysis of the genome of Campylobacter pylori with a rapid extraction method: evidence for considerable genomic variation. J Infect Dis. 1988 Mar;157(3):465–471. doi: 10.1093/infdis/157.3.465. [DOI] [PubMed] [Google Scholar]
  15. Mills S. D., Kurjanczyk L. A., Penner J. L. Antigenicity of Helicobacter pylori lipopolysaccharides. J Clin Microbiol. 1992 Dec;30(12):3175–3180. doi: 10.1128/jcm.30.12.3175-3180.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morgan D. R., Mathewson J. J., Freedman R., Kraft W. G. Evaluation of a selective enrichment technique for the isolation of Campylobacter pylori. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):303–306. doi: 10.1016/0378-1097(90)90302-7. [DOI] [PubMed] [Google Scholar]
  17. Oudbier J. H., Langenberg W., Rauws E. A., Bruin-Mosch C. Genotypical variation of Campylobacter pylori from gastric mucosa. J Clin Microbiol. 1990 Mar;28(3):559–565. doi: 10.1128/jcm.28.3.559-565.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Owen R. J., Bell G. D., Desai M., Moreno M., Gant P. W., Jones P. H., Linton D. Biotype and molecular fingerprints of metronidazole-resistant strains of Helicobacter pylori from antral gastric mucosa. J Med Microbiol. 1993 Jan;38(1):6–12. doi: 10.1099/00222615-38-1-6. [DOI] [PubMed] [Google Scholar]
  19. Owen R. J., Bickley J., Costas M., Morgan D. R. Genomic variation in Helicobacter pylori: application to identification of strains. Scand J Gastroenterol Suppl. 1991;181:43–50. doi: 10.3109/00365529109093207. [DOI] [PubMed] [Google Scholar]
  20. Owen R. J., Bickley J., Lastovica A., Dunn J. P., Borman P., Hunton C. Ribosomal RNA gene patterns of Helicobacter pylori from surgical patients with healed and recurrent peptic ulcers. Epidemiol Infect. 1992 Feb;108(1):39–50. doi: 10.1017/s0950268800049487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Owen R. J., Bickley J., Moreno M., Costas M., Morgan D. R. Biotype and macromolecular profiles of cytotoxin-producing strains of Helicobacter pylori from antral gastric mucosa. FEMS Microbiol Lett. 1991 Apr 15;63(2-3):199–204. doi: 10.1016/0378-1097(91)90086-p. [DOI] [PubMed] [Google Scholar]
  22. Owen R. J., Desai M., Figura N., Bayeli P. F., Di Gregorio L., Russi M., Musmanno R. A. Comparisons between degree of histological gastritis and DNA fingerprints, cytotoxicity and adhesivity of Helicobacter pylori from different gastric sites. Eur J Epidemiol. 1993 May;9(3):315–321. doi: 10.1007/BF00146270. [DOI] [PubMed] [Google Scholar]
  23. Owen R. J., Figura N., Moreno M. Biotypes and DNA fingerprints of cytotoxigenic Helicobacter pylori from patients with gastritis and peptic ulceration in Italy. Eur J Epidemiol. 1992 Jan;8(1):15–21. doi: 10.1007/BF02427386. [DOI] [PubMed] [Google Scholar]
  24. Owen R. J., Fraser J., Costas M., Morgan D., Morgan D. R. Signature patterns of DNA restriction fragments of Helicobacter pylori before and after treatment. J Clin Pathol. 1990 Aug;43(8):646–649. doi: 10.1136/jcp.43.8.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Owen R. J., Hunton C., Bickley J., Moreno M., Linton D. Ribosomal RNA gene restriction patterns of Helicobacter pylori: analysis and appraisal of Hae III digests as a molecular typing system. Epidemiol Infect. 1992 Aug;109(1):35–47. [PMC free article] [PubMed] [Google Scholar]
  26. Owen R. J., Martin S. R., Borman P. Rapid urea hydrolysis by gastric Campylobacters. Lancet. 1985 Jan 12;1(8420):111–111. doi: 10.1016/s0140-6736(85)92004-5. [DOI] [PubMed] [Google Scholar]
  27. Prewett E. J., Bickley J., Owen R. J., Pounder R. E. DNA patterns of Helicobacter pylori isolated from gastric antrum, body, and duodenum. Gastroenterology. 1992 Mar;102(3):829–833. doi: 10.1016/0016-5085(92)90165-u. [DOI] [PubMed] [Google Scholar]
  28. Rauws E. A., Tytgat G. N. Cure of duodenal ulcer associated with eradication of Helicobacter pylori. Lancet. 1990 May 26;335(8700):1233–1235. doi: 10.1016/0140-6736(90)91301-p. [DOI] [PubMed] [Google Scholar]
  29. Salvi R. J., Ahroon W., Saunders S. S., Arnold S. A. Evoked potentials: computer-automated threshold-tracking procedure using an objective detection criterion. Ear Hear. 1987 Jun;8(3):151–156. [PubMed] [Google Scholar]
  30. Simor A. E., Shames B., Drumm B., Sherman P., Low D. E., Penner J. L. Typing of Campylobacter pylori by bacterial DNA restriction endonuclease analysis and determination of plasmid profile. J Clin Microbiol. 1990 Jan;28(1):83–86. doi: 10.1128/jcm.28.1.83-86.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Taylor D. N., Blaser M. J. The epidemiology of Helicobacter pylori infection. Epidemiol Rev. 1991;13:42–59. doi: 10.1093/oxfordjournals.epirev.a036078. [DOI] [PubMed] [Google Scholar]
  32. Tee W., Lambert J., Smallwood R., Schembri M., Ross B. C., Dwyer B. Ribotyping of Helicobacter pylori from clinical specimens. J Clin Microbiol. 1992 Jun;30(6):1562–1567. doi: 10.1128/jcm.30.6.1562-1567.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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