Advertisement

Detection ofHelicobacter pylori in gastric biopsy tissue by polymerase chain reaction

  • J. T. Wang
  • J. T. Lin
  • J. C. Sheu
  • J. C. Yang
  • D. S. Chen
  • T. H. Wang
Notes

Abstract

To evaluate the sensitivity of a polymerase chain reaction (PCR) assay using nested primers in detectingHelicobacter pylori, gastric tissue biopsy specimens were collected on endoscopy from 17 patients with a duodenal ulcer. DNA was extracted by phenol/chloroform treatment or boiling in water, and then subjected to a nested PCR using two primer pairs from the urease gene ofHelicobacter pylori. Fourteen of the 17 patients were positive forHelicobacter pylori using DNA samples extracted by either method. The PCR results correlated well with the results of an enzyme immunoassay to detect IgG antibody. However, there were two culture negative patients. The three PCR negative patients were both culture negative and serologically negative. DNA from 9 of the 14 patients was randomly selected and subjected to semiquantification by serial dilutions, and then PCR. The results showed that phenol/chloroform extraction yielded 10–1000 times more DNA than the boiling method. It is concluded that the PCR assay is a rapid and sensitive method for detectingHelicobacter pylori, and that phenol/chloroform extraction is superior to simple boiling in obtaining DNA samples for PCR.

Keywords

Polymerase Chain Reaction Duodenal Ulcer Polymerase Chain Reaction Assay Negative Patient Nest Polymerase Chain Reaction 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Buck GE Campylobacter pylori and gastroduodenal disease. Clinical Microbiology Reviews 1990, 3: 1–12.Google Scholar
  2. 2.
    Graham DY Campylobacter pylori and peptic ulcer disease. Gastroenterology 1989, 96: 615–625.Google Scholar
  3. 3.
    Coudron PE, Kirby DF Comparison of rapid urease tests, staining techniques, and growth on different solid media for detection ofCampylobacter pylori. Journal of Clinical Microbiology 1989, 27: 1527–1530.Google Scholar
  4. 4.
    Evans DJ, Evans DG, Graham DY, Klein PD A sensitive and specific serologic test for detection ofCampylobacter pylori infections. Gastroenterology 1989, 96: 1004–1008.Google Scholar
  5. 5.
    Goodwin CS, Blincow ED, Warren JR, Waters TE, Sanderson CR, Easton L Evaluation of cultural techniques for isolatingCampylobacter pylori from endoscopic biopsies of gastric mucosa. Journal of Clinical Pathology 1985, 38: 1127–1131.Google Scholar
  6. 6.
    Marshall BJ, Surveyor I Carbon-14 urea breath test for the diagnosis ofCampylobacter pylori associated with gastritis. Journal of Nuclear Medicine 1988, 29: 11–16.Google Scholar
  7. 7.
    Mulrooney SB, Hausinger RP Sequence of theKlebsiella aerogenes urease genes and evidence for accessory proteins facilitating nickel incorporation. Journal of Bacteriology 1990, 172: 5837–5843.Google Scholar
  8. 8.
    Simor AE, Cooter NB, Low DE Comparison of four strains and a urease test for rapid detection ofCampylobacter pylori in gastric biopsies. European Journal of Clinical Microbiology and Infectious Diseases 1990, 9: 350–352.Google Scholar
  9. 9.
    Morotomi M, Hoshina S, Green P, Neu HC, LoGerfo P, Watanabe I, Mutai M, Weinstein IB Oligonucleotide probe for detection and identification ofCampylobacter pylori. Journal of Clinical Microbiology 1989, 27: 2652–2655.Google Scholar
  10. 10.
    Vandenberg FM, Zijlmans H, Langenberg W, Rauws E, Schipper M Detection ofCampylobacter pylori in stomach tissue by DNA in situ hybridization. Journal of Clinical Pathology 1989, 42: 995–1000.Google Scholar
  11. 11.
    Wetherall BL, McDonald PJ, Johnson AM Detection ofCampylobacter pylori DNA by hybridization with non-radioactive probes in comparison with a32p-labelled probe. Journal of Medical Microbiology 1988, 26: 257–263.Google Scholar
  12. 12.
    Valentine JL, Arthur RR, Mobley HLT, Dick JD Detection ofHelicobacter pylori by using the polymerase chain reaction. Journal of Clinical Microbiology 1991, 29: 689–695.Google Scholar
  13. 13.
    Clayton CL, Cleanthous H, Coates PJ, Morgan DD, Tabaqchali S Sensitive detection ofHelicobacter pylori by using polymerase chain reaction. Journal of Clinical Microbiology 1992, 30: 192–200.Google Scholar
  14. 14.
    Hammar M, Tyszkiewicz T, Wadstrom T, O'Toole PW Rapid detection ofHelicobacter pylori in gastric biopsy material by polymerase chain reaction. Journal of Clinical Microbiology 1992, 30: 54–58.Google Scholar
  15. 15.
    Clayton CL, Pallen MJ, Kleanthous H, Wren BW, Tabaqchali S Nucleotide sequence of two genes fromHelicobacter pylori encoding for urease subunits. Nucleic Acids Research 1990, 18: 362.Google Scholar
  16. 16.
    Wang JT, Wang TH, Sheu JC, Shih LN, Lin JT, Chen DS Detection of hepatitis B virus DNA by polymerase chain reaction in plasma of volunteer blood donors negative for hepatitis B surface antigen. Journal of Infectious Diseases 1991, 163: 397–399.Google Scholar
  17. 17.
    Wang JT, Wang TH, Sheu JC, Lin SM, Lin JT, Chen DS Effect of anticoagulants and storage of blood samples on efficacy of the polymerase chain reaction assay for hepatitis C virus. Journal of Clinical Microbiology 1992, 30: 750–753.Google Scholar
  18. 18.
    Kwok S, Higuchi R Avoiding false positives with PCR. Nature 1989, 339: 237–238.Google Scholar
  19. 19.
    Graham DY, Lew GM, Klein PD, Evans DG, Evans DJ, Saeed ZA, Malaty HM Effect of treatment ofHelicobacter pylori infection on the long-term recurrence of gastric or duodenal ulcer. A randomized, controlled study. Annals of Internal Medicine 1992, 116: 705–708.Google Scholar
  20. 20.
    Rauws EAJ, Royen EAV, Langenberg W, Woensel JV, Vrij AA, Tytgat GN C-14-urea breath test inCampylobacter pylori gastritis. Gut 1989, 30: 798–803.Google Scholar
  21. 21.
    Schnell GA, Schubert TT Usefulness of culture, histology and urease testing in the detection ofCampylobacter pylori in gastric biopsies. American Journal of Gastroenterology 1989, 84: 133–137.Google Scholar
  22. 22.
    Borromeo M, Lambert JR, Pinkard KJ Evaluation of “CLO-Test” to detectCampylobacter pyloridis in gastric mucosa. Journal of Clinical Pathology 1987, 40: 462–463.Google Scholar
  23. 23.
    Newell DG, Rathbone BJ The serodiagnosis ofHelicobacter pylori infection. Serodiagnosis and Immunotherapy of Infectious Diseases 1989, 3: 1–5.Google Scholar
  24. 24.
    Goodwin CS, Armstrong JA Microbiological aspects ofHelicobacter pylori (Campylobacter pylori). European Journal of Clinical Microbiology and Infectious Diseases 1990, 9: 1–13.Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1993

Authors and Affiliations

  • J. T. Wang
    • 1
  • J. T. Lin
    • 1
  • J. C. Sheu
    • 1
  • J. C. Yang
    • 1
  • D. S. Chen
    • 1
  • T. H. Wang
    • 1
  1. 1.Department of Internal MedicineNational Taiwan University HospitalTaipeiTaiwan

Personalised recommendations