Reverse transcription polymerase chain reaction in the diagnosis of Helicobacter pylori infection in Finnish children

Background: The purpose of this study was to design a simplified polymerase chain reaction (PCR) technique for the detection of Helicobacter pylori an d to compare it with conventional diagnostic methods-culture and histology of gastric biopsy specimens. In addition, the capability of this technique to detect H, pylori in the gastric mucosal biopsies of originally H. pylori -negative children with gastritis or recurrent abdominal pain was investiga ted. Methods: Reverse transcriptase polymerase chain reaction (RT-PCR) using pol ymerase from Thermus thermophilus was applied to detect H. pylori 16S rRNA. Twenty-five children H. pylori-positive by culture and/or histology were u sed as positive control subjects. Sixteen healthy H. pylori-negative childr en served as negative control subjects. Biopsy specimens from gastric antru m and corpus from 81 children were examined by RT-PCR. Altogether, 30 had h istologic gastritis and 51 had nonspecific abdominal pain only, with no dis ease in histologic specimens. Histology and culture of N. pylori were negat ive in both patient groups. Results: Reverse transcription-polymerase chain reaction detected 24 of 25 tissue-positive and 0 of 16 tissue-negative cases, indicating 96% sensitivi ty and 100% specificity for the test. None of the culturally and histologic ally H, pylori-negative samples showed H, pylori colonization when analyzed by RT-PCR. Conclusions: RT-PCR using Thermus thermophilus polymerase is a fast and sim ple means of detecting H. pylori in gastric biopsy specimens. It is at leas t as specific and sensitive as conventional methods. In pediatric patients it may be necessary to take mon than two biopsy specimens to increase sensi tivity in cases of local or patchy colonization.