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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Diouf
A
A
UFR d'odontologie, laboratoire parodontites et maladies générales LU46, département de santé publique, université Paul-Sabatier, 3, chemin des Maraîchers, 31062 Toulouse cedex 04, France. dioufa2005@yahoo.fr
Martinez-Gomis
J
J
Miquel
M
M
Quesada
M
M
Lario
S
S
Sixou
M
M
D001426:Bacterial Proteins / Q000235:genetics; D003199:Computer Systems; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D005756:Gastritis / Q000382:microbiology* / Q000473:pathology; D016481:Helicobacter Infections / Q000382:microbiology*; D016480:Helicobacter pylori / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D009055:Mouth / Q000382:microbiology; D010733:Phosphoglucomutase / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D021521:Ribotyping / Q000379:methods; D012680:Sensitivity and Specificity; D013270:Stomach / Q000382:microbiology
D001426:Bacterial Proteins; D015342:DNA Probes; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C097631:phosphoglucosamine mutase; D010733:Phosphoglucomutase
Comparaison de quatre paires d'amorces différentes dans la détection d'Helicobacter pylori dans les biopsies gastriques et les prélèvements oraux.