Pitfalls of UBT, SAT, and Nested PCR Reliability for Diagnosing Helicobacter pylori

Helicobacter pylori is the leading cause of chronic gastrointestinal tract diseases, with a worldwide prevalence of around 50%. For identification in medical practice, non-invasive methods such as the immunochromatographic test for antigen in stool (SAT) and the urease breath test (UBT) are widely used. Recently, we developed a highly sensitive and specific nested PCR (NPCR) that involves two amplification reactions and uses primers designed to target the variable regions of the 16S rRNA gene to amplify a short 148 bp amplicon. The aim of this study was to compare two classical methods, SAT and UBT, with the 148 bp amplicon NPCR. We examined samples from 137 volunteers, and found 46 positives with NPCR using stool samples, 34 with UBT, and only 24 with SAT. H. pylori origin of the 148 bp amplicons was confirmed by sequencing. NPCR had the highest detection rate in this cohort, suggesting that a portion of the population may be misdiagnosed, particularly by SAT. Due to the cost and simple performance in practice, SAT is a method of choice for initial screening. However, in cases of negative results and persistent digestive problems, we recommend the more sensitive UBT. NPCR may be a useful complementary method, especially in discordant or clinically suspicious cases.