Development of a PCR-based technique for detection of Helicobacter pylori |
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| Authors: | A-CE Thoreson MB Borre LP Andersen L Elsborg S Holck P Conway J Henrichsen J Vuust KA Krogfelt |
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| Institution: | Department of Bacteriology, Statens Seruminstitut, Artillerivej 5, 2300 Copenhagen-S, Denmark;Laboratory of Molecular Biology, Department of Infection-Immunology, Statens Seruminstitut, Artillerivej 5, 2300 Copenhagen-S, Denmark;Department of General and Marine Microbiology, University of Gothenburg, Laboratory of Lundberg, Medicinaregatan 9C, 413 90 Göteborg, Sweden;Department of Clinical Microbiology, Hillerød Hospital, DK-3400 Hillerød, Denmark;Department of Medicine B, Hillerød Hospital, DK-3400 Hillerød, Denmark;Department of Pathology, Hillerød Hospital, DK-3400 Hillerød, Denmark |
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| Abstract: | 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. |
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| Keywords: | Helicobacter pylori 16S rDNA-primer Polymerase chain reaction (PCR) Gastric human biopsy |
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