Development of a PCR-based technique for detection of Helicobacter pylori

A. C.E. Thoreson, M. B. Borre, L. P. Andersen, L. Elsborg, S. Holck, P. Conway, J. Henrichsen, J. Vuust, K. A. Krogfelt*

*Corresponding author for this work

Research output: Contribution to journalJournal articlepeer-review

Abstract

A primer-set was designed for specific detection of genes that encode for 16S rRNA of Helicobacter pylori, using direct polymerase chain reaction (PCR). The primers were selected in the hypervariable regions, derived from a complete small subunit 16S rRNA sequence of the reference strain H. pylori CCUG 17874. The primer-set amplified a 537 base pair (bp) sequence specifically from chromosomal H. pylori DNA. Amplification of purified chromosomal H. pylori DNA was achieved at concentrations as low as 1 femto gram (fg), equivalent to 5 bacteria. Furthermore, as few as 1 lysed H. pylori cell was detected by this PCR technique. The specificity of the primers was 100%, since purified chromosomal DNA was detected from all 32 various H. pylori isolates, whereas no other bacteria species were detected, whether related to Helicobacter or not. The 16S rDNA primers successfully detected H. pylori in antral biopsy specimens collected from infected patients.

Original languageEnglish
JournalFEMS Immunology and Medical Microbiology
Volume10
Issue number3-4
Pages (from-to)325-333
Number of pages9
ISSN0928-8244
DOIs
Publication statusPublished - Feb 1995
Externally publishedYes

Keywords

  • 16S rDNA-primer
  • Gastric human biopsy
  • Helicobacter pylori
  • Polymerase chain reaction (PCR)

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