Real-time PCR improves helicobacter pylori detection in patients with peptic ulcer bleeding

María José Ramírez-Lázaro, Sergio Lario, Alex Casalots, Esther Sanfeliu, Loreto Boix, Pilar García-Iglesias, Jordi Sánchez-Delgado, Antònia Montserrat, Maria Rosa Bella-Cueto, Marta Gallach, Isabel Sanfeliu, Ferran Segura, Xavier Calvet

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Abstract

Background and Aims: Histological and rapid urease tests to detect H. pylori in biopsy specimens obtained during peptic ulcer bleeding episodes (PUB) often produce false-negative results. We aimed to examine whether immunohistochemistry and real-time PCR can improve the sensitivity of these biopsies. Patients and Methods: We selected 52 histology-negative formalin-fixed paraffin-embedded biopsy specimens obtained during PUB episodes. Additional tests showed 10 were true negatives and 42 were false negatives. We also selected 17 histology-positive biopsy specimens obtained during PUB to use as controls. We performed immunohistochemistry staining and real-time PCR for 16S rRNA, ureA, and 23S rRNA for H. pylori genes on all specimens. Results: All controls were positive for H. pylori on all PCR assays and immunohistochemical staining. Regarding the 52 initially negative biopsies, all PCR tests were significantly more sensitive than immunohistochemical staining (p<0.01). Sensitivity and specificity were 55% and 80% for 16S rRNA PCR, 43% and 90% for ureA PCR, 41% and 80% for 23S rRNA PCR, and 7% and 100% for immunohistochemical staining, respectively. Combined analysis of PCR assays for two genes were significantly more sensitive than ureA or 23S rRNA PCR tests alone (p<0.05) and marginally better than 16S rRNA PCR alone. The best combination was 16S rRNA+ureA, with a sensitivity of 64% and a specificity of 80%. Conclusions: Real-time PCR improves the detection of H. pylori infection in histology-negative formalin-fixed paraffin-embedded biopsy samples obtained during PUB episodes. The low reported prevalence of H. pylori in PUB may be due to the failure of conventional tests to detect infection. © 2011 Ramírez-Lázaro et al.
Original languageEnglish
Article numbere20009
JournalPLoS ONE
Volume6
DOIs
Publication statusPublished - 1 Jan 2011

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    Ramírez-Lázaro, M. J., Lario, S., Casalots, A., Sanfeliu, E., Boix, L., García-Iglesias, P., Sánchez-Delgado, J., Montserrat, A., Bella-Cueto, M. R., Gallach, M., Sanfeliu, I., Segura, F., & Calvet, X. (2011). Real-time PCR improves helicobacter pylori detection in patients with peptic ulcer bleeding. PLoS ONE, 6, [e20009]. https://doi.org/10.1371/journal.pone.0020009