Sensitive detection of Helicobacter pylori by using polymerase chain reaction

C. L. Clayton, H. Kleanthous, P. J. Coates, D. D. Morgan, S. Tabaqchali

    Research output: Contribution to journalArticle

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    Abstract

    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.
    Original languageEnglish
    Pages (from-to)192-200
    Number of pages9
    JournalJournal of Clinical Microbiology
    Volume30
    Issue number1
    Publication statusPublished - 1992

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    Helicobacter pylori
    Polymerase Chain Reaction
    Urease
    Stomach
    Biopsy
    Agar Gel Electrophoresis
    DNA
    Paraffin
    Genes
    Microscopy

    Cite this

    Clayton, C. L., Kleanthous, H., Coates, P. J., Morgan, D. D., & Tabaqchali, S. (1992). Sensitive detection of Helicobacter pylori by using polymerase chain reaction. Journal of Clinical Microbiology, 30(1), 192-200.
    Clayton, C. L. ; Kleanthous, H. ; Coates, P. J. ; Morgan, D. D. ; Tabaqchali, S. / Sensitive detection of Helicobacter pylori by using polymerase chain reaction. In: Journal of Clinical Microbiology. 1992 ; Vol. 30, No. 1. pp. 192-200.
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    abstract = "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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    Clayton, CL, Kleanthous, H, Coates, PJ, Morgan, DD & Tabaqchali, S 1992, 'Sensitive detection of Helicobacter pylori by using polymerase chain reaction', Journal of Clinical Microbiology, vol. 30, no. 1, pp. 192-200.

    Sensitive detection of Helicobacter pylori by using polymerase chain reaction. / Clayton, C. L.; Kleanthous, H.; Coates, P. J.; Morgan, D. D.; Tabaqchali, S.

    In: Journal of Clinical Microbiology, Vol. 30, No. 1, 1992, p. 192-200.

    Research output: Contribution to journalArticle

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    AU - Clayton, C. L.

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    AU - Coates, P. J.

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    AU - Tabaqchali, S.

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    AB - 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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    Clayton CL, Kleanthous H, Coates PJ, Morgan DD, Tabaqchali S. Sensitive detection of Helicobacter pylori by using polymerase chain reaction. Journal of Clinical Microbiology. 1992;30(1):192-200.