Detection of Helicobacter pylori by using the polymerase chain reaction

J. L. Valentine, R. R. Arthur, H. L T Mobley, J. D. Dick

Research output: Contribution to journalArticle

Abstract

A 1.9-kb cloned fragment of chromosomal DNA randomly selected from a Helicobacter pylori cloned library was evaluated as a potential probe. The probe detected 19 of 19 H. pylori strains and yielded a specificity of 98.7% when tested against 306 other bacterial strains representing 32 different species. False-positive results with non-H. pylori strains were due to the presence of contaminating vector sequences. A polymerase chain reaction (PCR) assay was developed by using 20-base oligonucleotide primers homologous to a portion of the 1.9-kb fragment. The PCR assay amplified a 203-nucleotide-pair product which was analyzed by agarose gel electrophoresis and Southern hybridization by using a third 20-base 32P-labeled oligonucleotide complementary to a region of DNA between the primers. The PCR assay was 100% sensitive, detecting all 35 H. pylori strains tested, and did not amplify sequences in several closely related species. The assay was sensitive for as little as one copy of the cloned plasmid DNA or 100 H. pylori bacterial cells. To evaluate the PCR assay for clinical samples, gastric biopsy and aspirate specimens were tested by PCR, and the results were compared with those of microbiologic culture and histologic examination. In fresh biopsy specimens, H. pylori sequences were detected by PCR in 13 of 14 (93%) positive tissues and 0 of 19 negative tissues. In gastric aspirate specimens, 11 of 13 (85%) positive tissues were positive by PCR. H. pylori DNA was detected in 1 of 14 aspirate specimens negative by culture, histology, and PCR of the accompanying biopsy tissue. PCR is a rapid, accurate, and sensitive method for the detection of H. pylori.

Original languageEnglish (US)
Pages (from-to)689-695
Number of pages7
JournalJournal of Clinical Microbiology
Volume29
Issue number4
StatePublished - 1991

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Helicobacter pylori
Polymerase Chain Reaction
DNA Primers
Biopsy
Stomach
Agar Gel Electrophoresis
DNA
Pylorus
Oligonucleotides
Libraries
Histology
Plasmids
Nucleotides

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Valentine, J. L., Arthur, R. R., Mobley, H. L. T., & Dick, J. D. (1991). Detection of Helicobacter pylori by using the polymerase chain reaction. Journal of Clinical Microbiology, 29(4), 689-695.

Detection of Helicobacter pylori by using the polymerase chain reaction. / Valentine, J. L.; Arthur, R. R.; Mobley, H. L T; Dick, J. D.

In: Journal of Clinical Microbiology, Vol. 29, No. 4, 1991, p. 689-695.

Research output: Contribution to journalArticle

Valentine, JL, Arthur, RR, Mobley, HLT & Dick, JD 1991, 'Detection of Helicobacter pylori by using the polymerase chain reaction', Journal of Clinical Microbiology, vol. 29, no. 4, pp. 689-695.
Valentine JL, Arthur RR, Mobley HLT, Dick JD. Detection of Helicobacter pylori by using the polymerase chain reaction. Journal of Clinical Microbiology. 1991;29(4):689-695.
Valentine, J. L. ; Arthur, R. R. ; Mobley, H. L T ; Dick, J. D. / Detection of Helicobacter pylori by using the polymerase chain reaction. In: Journal of Clinical Microbiology. 1991 ; Vol. 29, No. 4. pp. 689-695.
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