Abstract

Background: Helicobacter pylori is the most prevalent chronic bacterial infection globally, acquired mostly during childhood. It is associated with chronic gastritis, peptic ulcer disease, and gastric cancer. Due to challenges in culturing H. pylori, diagnostic reference standards often rely on combining >= 2 non-culture, biopsy-based methods. Histology with Giemsa staining is widely used in clinical settings due to its low cost and reliable performance. Methods: This study evaluated the concordance between histology with Giemsa staining as the reference standard and other diagnostic methods, including the rapid urease test (RUT), ureA RT-PCR, 16S sequencing, and anti-H. pylori serum IgG. Positive percent of agreement (PPA), negative percent of agreement (NPA) and concordance kappa index were calculated. Results: A total of 120 patients (41 positive and 79 negative by Giemsa staining) were analyzed. Among the methods tested, RT-PCR for ureA showed the best performance (PPA = 94.7%, NPA = 98.6%, kappa = 0.939), while RUT underperformed compared with expectations (PPA = 65.9%, NPA = 97.5%, kappa = 0.681). Serology had the lowest performance (PPA = 53.7%, NPA = 96.1%, kappa = 0.548). Conclusions: The combination of histology with Giemsa staining and ureA RT-PCR achieved the highest detection rate and strongest agreement.