[Evaluation of a simple rapid polymerase chain reaction (PCR) technique for the diagnosis of Helicobacter pylori infection in childhood]

Abstract

Background: The purpose of this study was to evaluate the use of polymerase chain reaction (PCR) applied to the diagnosis Helicobacter pylori infection in the pediatric population, by means of a rapid and simple method of extraction and posterior detection by a colorimetric hybridation of amplified H. pylori DNA.

Methods: Fifty three gastric biopsies, obtained through upper gastrointestinal endoscopy from 53 patients with gastric pathology (45 recidivant abdominal pain and eight hematemesis), were processed from october 1995 to july 1996. Three non invasive tests were performed: detection of IgG by (Cobas) Core Anti-H. pylori EIA (Roche), breath test with 13C-urea and PCR of dental plaque, as well as four invasive ones: histologic study, culture into selective (Pylori Agar, bioMérieux) and non selective media (Columbia Agar with 5% sheep blood, bioMérieux), test of rapid urease and PCR of gastric biopsies. A 20% solution of Chelex 100 resin (DNA Extraction Reagen, Perkin Elmer) was used for DNA extraction, amplification was performed from gen ureA (Clayton, 1992) and amplified DNA was detected by colorimetric hybridation (PCR ELISA, Boehringer, Mannheim). Duration of the PCR process was: extraction 25 min, amplification two hours and detection three hours.

Results: Results of culture and PCR from gastric biopsies agreed in 84.3% of cases (27 positives, 16 negatives and two not determined). Two samples were positive by culture and negative by PCR, and were considered as PCR false negatives due to positivity of three or more other tests. Six samples were negative by culture and positive by PCR, being considered as culture false negatives due to positivity of three or more other tests. Sensitivity of PCR and culture was 94.2 and 82.8%. Specificity was 100% for both tests. None of the dental plaque samples was positive.

Conclusions: When invasive techniques are to be done for microbiologic diagnose of H. pylori, PCR increases the confirmation rate of infection; the present procedure enables daily routine work due to its simplicity and its short turnaround time.