Comparison of four different primer sets for detection of Helicobacter pylori in gastric biopsies and oral samples by using real-time PCR

The presence of Helicobacter pylori. in the oral cavity remains controversial and the most appropriate method for detection of oral H. pylori has yet to be established. The aim of the present study was to compare four different primer sets on the detection of H. pylori in gastric biopsies and oral s...

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Bibliographic Details
Published in:Pathologie biologie (Paris) 2009-02, Vol.57 (1), p.30-35
Main Authors: Diouf, A, Martinez-Gomis, J, Miquel, M, Quesada, M, Lario, S, Sixou, M
Format: Article
Language:fre
Subjects:
Bacterial Proteins - genetics
Computer Systems
DNA Probes
DNA, Bacterial - analysis
DNA, Ribosomal - analysis
Gastritis - microbiology
Gastritis - pathology
Helicobacter Infections - microbiology
Helicobacter pylori - enzymology
Helicobacter pylori - genetics
Helicobacter pylori - isolation & purification
Humans
Mouth - microbiology
Phosphoglucomutase - genetics
Polymerase Chain Reaction - methods
Ribotyping - methods
RNA, Bacterial - genetics
RNA, Ribosomal, 16S - genetics
Sensitivity and Specificity
Stomach - microbiology
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Summary:The presence of Helicobacter pylori. in the oral cavity remains controversial and the most appropriate method for detection of oral H. pylori has yet to be established. The aim of the present study was to compare four different primer sets on the detection of H. pylori in gastric biopsies and oral samples using real-time PCR. Gastric biopsy and oral samples were collected from eight patients with gastric symptoms. DNA from clinical samples was extracted and analyzed for the presence of H. pylori by real-time PCR (LightCycler 2.0) using four pairs of primers which targeted 16S rRNA (16S rRNA#295; 16S rRNA#120) or glmM (glmM#294; glmM#722) DNA genes. Three H. pylori strains and three clinical isolates served as reference. The specificities of the four primer pairs were examined for seven oral microorganisms and two Helicobacter non-pylori species. Primer pair 16S rRNA#120 showed an acceptable specificity and a high sensitivity. Primer pairs glmM#294 and glmM#722 demonstrated a high specificity but a low sensitivity and primer pair 16S rRNA#295 demonstrated a poor specificity but acceptable sensitivity. Four H. pylori positive gastric biopsies were demonstrated by culture, histology and real-time PCR with primer pairs 16S rRNA#295 or 16S rRNA#120. No H. pylori was detected in oral samples, either by culture or by real-time PCR. Of the four different primer pairs examined, 16S rRNA#120 was the most appropriate to detect H. pylori in clinical samples using real-time PCR.
ISSN:0369-8114
1768-3114
DOI:10.1016/j.patbio.2008.07.008