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J Clin Microbiol. Jan 1992; 30(1): 192–200.
PMCID: PMC265019

Sensitive detection of Helicobacter pylori by using polymerase chain reaction.


A polymerase chain reaction (PCR) for the specific detection of Helicobacter pylori was developed with a single primer pair derived from the nucleotide sequence of the urease A gene of H. pylori. We achieved specific amplification of a 411-bp DNA fragment in H. pylori. After 35 cycles of amplification, the product could be detected by agarose gel electrophoresis and contained conserved single HinfI and AluI restriction sites. This fragment was amplified in all 50 strains of H. pylori tested, but it was not detected in other bacterial species, showing the PCR assay to be 100% specific. PCR DNA amplification was able to detect as few as 10 H. pylori cells. PCR detected H. pylori in 15 of 23 clinical human gastric biopsy samples, whereas culturing and microscopy detected H. pylori in only 7 of the samples found to be positive by PCR. Additional primer pairs based on the urease genes enabled the detection of H. pylori in paraffin-embedded human gastric biopsy samples. The detection of H. pylori by PCR will enable both retrospective and prospective analyses of clinical samples, elucidating the role of this organism in gastroduodenal disease.

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  • Blanchard A. Ureaplasma urealyticum urease genes; use of a UGA tryptophan codon. Mol Microbiol. 1990 Apr;4(4):669–676. [PubMed]
  • Borromeo M, Lambert JR, Pinkard KJ. Evaluation of "CLO-test" to detect Campylobacter pyloridis in gastric mucosa. J Clin Pathol. 1987 Apr;40(4):462–463. [PMC free article] [PubMed]
  • Buck GE. Campylobacter pylori and gastroduodenal disease. Clin Microbiol Rev. 1990 Jan;3(1):1–12. [PMC free article] [PubMed]
  • Burstain JM, Grimprel E, Lukehart SA, Norgard MV, Radolf JD. Sensitive detection of Treponema pallidum by using the polymerase chain reaction. J Clin Microbiol. 1991 Jan;29(1):62–69. [PMC free article] [PubMed]
  • Claas HC, Wagenvoort JH, Niesters HG, Tio TT, Van Rijsoort-Vos JH, Quint WG. Diagnostic value of the polymerase chain reaction for Chlamydia detection as determined in a follow-up study. J Clin Microbiol. 1991 Jan;29(1):42–45. [PMC free article] [PubMed]
  • Clayton CL, Pallen MJ, Kleanthous H, Wren BW, Tabaqchali S. Nucleotide sequence of two genes from Helicobacter pylori encoding for urease subunits. Nucleic Acids Res. 1990 Jan 25;18(2):362–362. [PMC free article] [PubMed]
  • Clayton CL, Wren BW, Mullany P, Topping A, Tabaqchali S. Molecular cloning and expression of Campylobacter pylori species-specific antigens in Escherichia coli K-12. Infect Immun. 1989 Feb;57(2):623–629. [PMC free article] [PubMed]
  • Coates PJ, d'Ardenne AJ, Khan G, Kangro HO, Slavin G. Simplified procedures for applying the polymerase chain reaction to routinely fixed paraffin wax sections. J Clin Pathol. 1991 Feb;44(2):115–118. [PMC free article] [PubMed]
  • Coudron PE, Kirby DF. Comparison of rapid urease tests, staining techniques, and growth on different solid media for detection of Campylobacter pylori. J Clin Microbiol. 1989 Jul;27(7):1527–1530. [PMC free article] [PubMed]
  • Das SS, Bain LA, Karim QN, Coelho LG, Baron JH. Rapid diagnosis of Campylobacter pyloridis infection. J Clin Pathol. 1987 Jun;40(6):701–702. [PMC free article] [PubMed]
  • Eisenach KD, Cave MD, Bates JH, Crawford JT. Polymerase chain reaction amplification of a repetitive DNA sequence specific for Mycobacterium tuberculosis. J Infect Dis. 1990 May;161(5):977–981. [PubMed]
  • Engstrand L, Pählson C, Gustavsson S, Schwan A. Monoclonal antibodies for rapid identification of Campylobacter pyloridis. Lancet. 1986 Dec 13;2(8520):1402–1403. [PubMed]
  • Evans DJ, Jr, Evans DG, Graham DY, Klein PD. A sensitive and specific serologic test for detection of Campylobacter pylori infection. Gastroenterology. 1989 Apr;96(4):1004–1008. [PubMed]
  • Goodwin CS, Armstrong JA. Microbiological aspects of Helicobacter pylori (Campylobacter pylori). Eur J Clin Microbiol Infect Dis. 1990 Jan;9(1):1–13. [PubMed]
  • Goodwin CS, Blincow ED, Warren JR, Waters TE, Sanderson CR, Easton L. Evaluation of cultural techniques for isolating Campylobacter pyloridis from endoscopic biopsies of gastric mucosa. J Clin Pathol. 1985 Oct;38(10):1127–1131. [PMC free article] [PubMed]
  • Graham DY. Campylobacter pylori and peptic ulcer disease. Gastroenterology. 1989 Feb;96(2 Pt 2 Suppl):615–625. [PubMed]
  • Hance AJ, Grandchamp B, Lévy-Frébault V, Lecossier D, Rauzier J, Bocart D, Gicquel B. Detection and identification of mycobacteria by amplification of mycobacterial DNA. Mol Microbiol. 1989 Jul;3(7):843–849. [PubMed]
  • Jackson DP, Lewis FA, Taylor GR, Boylston AW, Quirke P. Tissue extraction of DNA and RNA and analysis by the polymerase chain reaction. J Clin Pathol. 1990 Jun;43(6):499–504. [PMC free article] [PubMed]
  • Jones BD, Mobley HL. Proteus mirabilis urease: nucleotide sequence determination and comparison with jack bean urease. J Bacteriol. 1989 Dec;171(12):6414–6422. [PMC free article] [PubMed]
  • Krajden S, Fuksa M, Anderson J, Kempston J, Boccia A, Petrea C, Babida C, Karmali M, Penner JL. Examination of human stomach biopsies, saliva, and dental plaque for Campylobacter pylori. J Clin Microbiol. 1989 Jun;27(6):1397–1398. [PMC free article] [PubMed]
  • Kwok S, Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. [PubMed]
  • Campylobacter-like organisms in the stomach of patients and healthy individuals. Lancet. 1984 Jun 16;1(8390):1348–1349. [PubMed]
  • Mahbubani MH, Bej AK, Miller R, Haff L, DiCesare J, Atlas RM. Detection of Legionella with polymerase chain reaction and gene probe methods. Mol Cell Probes. 1990 Jun;4(3):175–187. [PubMed]
  • Marshall BJ, Surveyor I. Carbon-14 urea breath test for the diagnosis of Campylobacter pylori associated gastritis. J Nucl Med. 1988 Jan;29(1):11–16. [PubMed]
  • Marshall BJ, Goodwin CS, Warren JR, Murray R, Blincow ED, Blackbourn SJ, Phillips M, Waters TE, Sanderson CR. Prospective double-blind trial of duodenal ulcer relapse after eradication of Campylobacter pylori. Lancet. 1988 Dec 24;2(8626-8627):1437–1442. [PubMed]
  • Morotomi M, Hoshina S, Green P, Neu HC, LoGerfo P, Watanabe I, Mutai M, Weinstein IB. Oligonucleotide probe for detection and identification of Campylobacter pylori. J Clin Microbiol. 1989 Dec;27(12):2652–2655. [PMC free article] [PubMed]
  • Mulrooney SB, Hausinger RP. Sequence of the Klebsiella aerogenes urease genes and evidence for accessory proteins facilitating nickel incorporation. J Bacteriol. 1990 Oct;172(10):5837–5843. [PMC free article] [PubMed]
  • Nedenskov-Sørensen P, Aase S, Bjørneklett A, Fausa O, Bukholm G. Sampling efficiency in the diagnosis of Helicobacter pylori infection and chronic active gastritis. J Clin Microbiol. 1991 Apr;29(4):672–675. [PMC free article] [PubMed]
  • Plikaytis BB, Gelber RH, Shinnick TM. Rapid and sensitive detection of Mycobacterium leprae using a nested-primer gene amplification assay. J Clin Microbiol. 1990 Sep;28(9):1913–1917. [PMC free article] [PubMed]
  • Rauws EA, Langenberg W, Houthoff HJ, Zanen HC, Tytgat GN. Campylobacter pyloridis-associated chronic active antral gastritis. A prospective study of its prevalence and the effects of antibacterial and antiulcer treatment. Gastroenterology. 1988 Jan;94(1):33–40. [PubMed]
  • Rauws EA, Royen EA, Langenberg W, Woensel JV, Vrij AA, Tytgat GN. 14C-urea breath test in C pylori gastritis. Gut. 1989 Jun;30(6):798–803. [PMC free article] [PubMed]
  • Rogers BB, Alpert LC, Hine EA, Buffone GJ. Analysis of DNA in fresh and fixed tissue by the polymerase chain reaction. Am J Pathol. 1990 Mar;136(3):541–548. [PMC free article] [PubMed]
  • Rosa PA, Schwan TG. A specific and sensitive assay for the Lyme disease spirochete Borrelia burgdorferi using the polymerase chain reaction. J Infect Dis. 1989 Dec;160(6):1018–1029. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. [PubMed]
  • Schaber E, Umlauft F, Stöffler G, Aigner F, Paulweber B, Sandhofer F. Indirect immunofluorescence test and enzyme-linked immunosorbent assay for detection of Campylobacter pylori. J Clin Microbiol. 1989 Feb;27(2):327–330. [PMC free article] [PubMed]
  • Schnell GA, Schubert TT. Usefulness of culture, histology, and urease testing in the detection of Campylobacter pylori. Am J Gastroenterol. 1989 Feb;84(2):133–137. [PubMed]
  • Shibata D, Martin WJ, Arnheim N. Analysis of DNA sequences in forty-year-old paraffin-embedded thin-tissue sections: a bridge between molecular biology and classical histology. Cancer Res. 1988 Aug 15;48(16):4564–4566. [PubMed]
  • Simor AE, Cooter NB, Low DE. Comparison of four stains and a urease test for rapid detection of Helicobacter pylori in gastric biopsies. Eur J Clin Microbiol Infect Dis. 1990 May;9(5):350–352. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Starnbach MN, Falkow S, Tompkins LS. Species-specific detection of Legionella pneumophila in water by DNA amplification and hybridization. J Clin Microbiol. 1989 Jun;27(6):1257–1261. [PMC free article] [PubMed]
  • Thillainayagam AV, Arvind AS, Cook RS, Harrison IG, Tabaqchali S, Farthing MJ. Diagnostic efficiency of an ultrarapid endoscopy room test for Helicobacter pylori. Gut. 1991 May;32(5):467–469. [PMC free article] [PubMed]
  • Vaira D, Holton J, Cairns S, Falzon M, Salmon P. Four hour rapid urease test (RUT) for detecting Campylobacter pylori: is it reliable enough to start treatment? J Clin Pathol. 1988 Mar;41(3):355–356. [PMC free article] [PubMed]
  • Valentine JL, Arthur RR, Mobley HL, Dick JD. Detection of Helicobacter pylori by using the polymerase chain reaction. J Clin Microbiol. 1991 Apr;29(4):689–695. [PMC free article] [PubMed]
  • West AP, Millar MR, Tompkins DS. Survival of Helicobacter pylori in water and saline. J Clin Pathol. 1990 Jul;43(7):609–609. [PMC free article] [PubMed]
  • Wetherall BL, McDonald PJ, Johnson AM. Detection of Campylobacter pylori DNA by hybridisation with non-radioactive probes in comparison with a 32P-labelled probe. J Med Microbiol. 1988 Aug;26(4):257–263. [PubMed]

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