Unreliability of results of PCR detection of Helicobacter pylori in clinical or environmental samples.
The aim of this study was to compare published Helicobacter pylori primer pairs for their ability to reliably detect H. pylori in gastric biopsy specimens and salivary samples. Detection limits of the 26 PCR primer pairs previously described for detection of H. pylori DNA in clinical samples were determined. Sensitivity and specificity were determined using primers with detection limits of <100 CFU/ml using 50 H. pylori-positive and -negative (by concordance by culture and histology) coded gastric biopsy specimens. These results were then confirmed with gastric biopsy specimens and saliva from patients with confirmed H. pylori status. Five of the twenty-six previously reported primer pairs (HP64-f/HP64-r, HP1/HP2, EHC-U/EHC-L, VAG-F/VAG-R, and ICT37/ICT38) had detection limits of <100 CFU/ml in the presence of gastric tissue. None had 100% specificity or sensitivity; all produced false-positive results. The HP64-f/HP64-r for ureA and HP1/HP2 for 16S rRNA individually had sensitivities and specificities of >90% with gastric biopsy specimens. No combinations of primer pairs improved the results. Using these five primer pairs, 54% of the positive saliva samples were determined to be false positive; both the HP64-f/HP64-r and the HP1/HP2 sets produced false positives with saliva. We conclude that clinicians should not rely on results using current PCR primers alone to decide the H. pylori status of an individual patient or as a basis for treatment decisions. The results of studies based on PCR identification of H. pylori in environmental samples should be viewed with caution. Possibly, specific primers sets can be identified based on the presence of multiple putative virulence factor genes.
Sugimoto
Mitsushige
M
Department of Medicine, Michael E. DeBakey Veterans Affairs Medical Center, Houston, Texas 77030, USA.
Wu
Jeng-Yih
JY
Abudayyeh
Suhaib
S
Hoffman
Jill
J
Brahem
Hajer
H
Al-Khatib
Khaldun
K
Yamaoka
Yoshio
Y
Graham
David Y
DY
D017931:DNA Primers / Q000235:genetics; D004783:Environmental Microbiology; D005189:False Positive Reactions; D005753:Gastric Mucosa / Q000382:microbiology; D016481:Helicobacter Infections / Q000175:diagnosis / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012463:Saliva / Q000382:microbiology; D012680:Sensitivity and Specificity
D017931:DNA Primers