Toward a non-invasive diagnostic tool for Helicobacter pylori ‒ insights from ELISA-based biomarker profiling
DOI:
https://doi.org/10.15584/ejcem.2026.1.15Keywords:
anti-CagA IgG, biomarker panel, ELISA, Helicobacter pylori, histopathology, non-invasive diagnosisAbstract
Introduction and aim. This study aimed to evaluate the diagnostic performance of a novel ELISA-based panel of virulence-associated antibodies (anti-CagA, anti-UreB, and anti-HpNAP IgG) for early detection of Helicobacter pylori infection.
Material and methods. In this cross-sectional study of 40 dyspeptic patients, ELISA results were compared with histopathology and stool antigen testing as reference standards. Diagnostic accuracy was assessed using receiver operating characteristic (ROC) curve analysis, and predictors were evaluated through logistic regression.
Results. Anti-CagA IgG achieved the highest diagnostic performance (AUC=0.95; sensitivity=90.9%; specificity=94.4%), followed by anti-UreB (AUC=0.92) and anti-HpNAP (AUC=0.89). The combined biomarker model reached an AUC of 0.97, demonstrating strong correlation with both infection status and symptom severity. Agreement between stool antigen testing and histopathology was high (κ=0.80).
Conclusion. This study provides the first regional validation of a standardized three-marker ELISA panel that demonstrated high accuracy as a non-invasive diagnostic approach for early H. pylori detection, offering a cost-effective tool for use in resource-limited settings.
Downloads
References
Shirani M, Pakzad R, Haddadi MH, et al. The global prevalence of gastric cancer in Helicobacter pylori-infected individuals: a systematic review and meta-analysis. BMC Infect Dis. 2023;23(1):543. doi:10.1186/s12879-023-08504-5
Kumar N. Status of Helicobacter pylori in gut microbiome and precision medicine. In: Precision Medicine and Human Health. Bentham Science Publishers; 2024:153-176. doi:10.2174/97898152235831240101
Chivu RF, Bobirca F, Melesteu I, Patrascu T. The role of Helicobacter pylori infection in the development of gastric cancer: review of the literature. Chirurgia (Bucur). 2024;119(eCollection):1-10. doi:10.21614/chirurgia.119.eC.2971
Jeon HJ, Choi HS. Various diagnostic methods for Helicobacter pylori infection. Korean J Med. 2024;99(2):104-110. doi:10.3904/kjm.2024.99.2.104
Lestari HM, Gustinanda J, Tejaputri NA, et al. Stool antigen examination as a diagnostic tool for dyspeptic patients caused by Helicobacter pylori infection: a case-based literature review. Open Access Maced J Med Sci. 2021;9(F):305-309. doi:10.3889/oamjms.2021.5568
Hussein RA, Al-Ouqaili MT, Majeed YH. Detection of Helicobacter pylori infection by invasive and non-invasive techniques in patients with gastrointestinal diseases from Iraq: a validation study. PLoS One. 2021;16(8):e0256393. doi:10.1371/journal.pone.0256393
Li Z-X, Bronny K, Formichella L, et al. A multiserological line assay to discriminate current from past Helicobacter pylori infection. Clin Microbiol Infect. 2024;30(1):114-121. doi:10.1016/j.cmi.2023.10.006
Fu H-W. Helicobacter pylori neutrophil-activating protein: from molecular pathogenesis to clinical applications. World J Gastroenterol. 2014;20(18):5294-5301. doi:10.3748/wjg.v20.i18.5294
Mahmoodzadeh AS, Moazamian E, Shamsedin SA, Kaydani GA. Prevalence of virulence genes and antigen pattern in Helicobacter pylori-infected patients and inflammatory cytokine levels compared with non-infected individuals. Jundishapur J Microbiol. 2022;15(2):e121144. doi:10.5812/jjm-121144
Hussein RA, Al-Ouqaili MT, Majeed YH. Detection of clarithromycin resistance and 23S rRNA point mutations in Helicobacter pylori: phenotypic and molecular methods. Saudi J Biol Sci. 2022;29(1):513-520. doi:10.1016/j.sjbs.2021.09.024
Ansari S, Yamaoka Y. Helicobacter pylori virulence factor CagA-mediated gastric pathogenicity. Int J Mol Sci. 2020;21(19):7430. doi:10.3390/ijms21197430
Atherton J. CagA, the cag pathogenicity island and Helicobacter pylori virulence. Gut. 1999;44(3):307-308. doi:10.1136/gut.44.3.307
Ilie M, Dascălu L, Chifiriuc C, et al. Correlation of anti-Helicobacter pylori CagA IgG with resistance to first-line treatment, bleeding gastroduodenal ulcers and gastric cancer. Roum Arch Microbiol Immunol. 2011;70(3):101-104.
Loffeld RJ, Werdmuller BF, Kusters JG, Kuipers EJ. IgG antibody titer against Helicobacter pylori correlates with CagA-positive strains. FEMS Immunol Med Microbiol. 2000;28(2):139-141. doi:10.1111/j.1574-695X.2000.tb01468.x
Codolo G, Coletta S, D’Elios MM, de Bernard M. HP-NAP of Helicobacter pylori: the power of immunomodulation. Front Immunol. 2022;13:944139. doi:10.3389/fimmu.2022.944139
Fu H-W, Lai Y-C. The role of Helicobacter pylori neutrophil-activating protein in pathogenesis and therapeutic development. Int J Mol Sci. 2022;24(1):91. doi:10.3390/ijms24010091
Wen Y, Feng J, Scott DR, Marcus EA, Sachs G. A cis-encoded antisense RNA controls expression of ureB in Helicobacter pylori. J Bacteriol. 2011;193(1):40-51. doi:10.1128/JB.00800-10
Ge R-G, Wang D-X, Hao M-C, Sun X-S. Nickel trafficking system responsible for urease maturation in Helicobacter pylori. World J Gastroenterol. 2013;19(45):8211-8217. doi:10.3748/wjg.v19.i45.8211
Schauer K, Muller C, Carriere M, et al. The Helicobacter pylori GroES cochaperonin HspA functions as a specialized nickel chaperone. J Bacteriol. 2010;192(5):1231-1237. doi:10.1128/JB.01216-09
Huifang G, Wenhong Z, Dongqing W, et al. Rapid screening of dominant B-cell epitopes of UreB by fluorescence polarization assay. Chin J Biochem Mol Biol. 2006;33(1):83-86. doi:10.1186/1556-276x-8-118
Peng J, Xie J, Liu D, et al. Impact of Helicobacter pylori colonization density and depth on gastritis severity. Ann Clin Microbiol Antimicrob. 2024;23(1):4. doi:10.1186/s12941-024-00666-7
Filomena A, Guenther A, Planatscher H, et al. Performance of a multiplex serological Helicobacter pylori assay on a microfluidic platform. Proteomes. 2017;5(4):24. doi:10.3390/proteomes5040024
Duquesne A, Falcón R, Galindo B, et al. Diagnostic accuracy for Helicobacter pylori infection among dyspeptic adults in Cuban primary care. Microorganisms. 2023;11(4):997. doi:10.3390/microorganisms11040997
Seo J-H, Lim CW, Park JS, et al. Correlations between CagA antigen and serum anti-Helicobacter pylori IgG and IgA in children. J Korean Med Sci. 2016;31(3):417-423. doi:10.3346/jkms.2016.31.3.417
Yu J-H, Zhao Y, Wang X-F, Xu Y-C. Evaluation of anti-Helicobacter pylori IgG for detection of infection in different populations. Diagnostics. 2022;12(5):1214. doi:10.3390/diagnostics12051214
Shiota S, Yamaoka Y. Biomarkers for Helicobacter pylori infection and gastroduodenal disease. Biomark Med. 2014;8(9):1127-1137. doi:10.2217/bmm.14.72
Han F-C, Li X-J, Jiang H, et al. Detection of Helicobacter pylori antibody profile in serum by protein array. World J Gastroenterol. 2006;12(25):4044-4049. doi:10.3748/wjg.v12.i25.4044
Hussein RA, Al-Ouqaili MT, Majeed YH. Association between alcohol, smoking, and Helicobacter pylori infection in Iraqi patients. J Emerg Med Trauma Acute Care. 2022;2022(6):12. doi:10.5339/jemtac.2022.aimco.12
Parsonnet J, Friedman G, Orentreich N, Vogelman H. Risk for gastric cancer in CagA-positive vs CagA-negative Helicobacter pylori infection. Gut. 1997;40(3):297-301. doi:10.1136/gut.40.3.297
Nomura AM, Lee J, Stemmermann GN, Nomura RY, Perez-Perez GI, Blaser MJ. Helicobacter pylori CagA seropositivity and gastric carcinoma risk in Japanese Americans. J Infect Dis. 2002;186(8):1138-1144. doi:10.1086/343808
Dudek I, Forma A, Hamerska J, et al. CagA virulence and epithelial–mesenchymal transition in gastric cancer. J Educ Health Sport. 2022;12(4):62-75. doi:10.12775/JEHS.2022.12.04.005
Wang H, Zhao M, Shi F, et al. Signaling pathways induced by Helicobacter pylori CagA. Front Cell Infect Microbiol. 2023;13:1062803. doi:10.3389/fcimb.2023.1062803
Covacci A, Censini S, Bugnoli M, et al. Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori. Proc Natl Acad Sci U S A. 1993;90(12):5791-5795. doi:10.1073/pnas.90.12.5791
Marzhoseyni Z, Mousavi MJ, Ghotloo S. Helicobacter pylori antigens as immunomodulators. Helicobacter. 2024;29(1):e13058. doi:10.1111/hel.13058
Carosella N, Brock KP, Zambelli B, et al. Functional contacts for urease activation in Helicobacter pylori. Front Chem Biol. 2023;2:1243564. doi:10.3389/fchbi.2023.1243564
Beswick EJ, Pinchuk IV, Minch K, et al. Urease B subunit binds CD74 and induces NF-κB and IL-8 in gastric epithelial cells. Infect Immun. 2006;74(2):1148-1155. doi:10.1128/IAI.74.2.1148-1155.2006
Kolopaking MS. Urease, gastric bacteria and gastritis. Acta Med Indones. 2022;54(1):1-1.
Wen S-H, Hong Z-W, Chen C-C, Chang H-W, Fu H-W. HP-NAP activates TLR2 inducing IL-8 secretion. Int J Mol Sci. 2021;22(21):11560. doi:10.3390/ijms222111560
Ji X, Sun Z, Wu H, et al. Stronger dysregulation by East Asian-type CagA on intracellular signaling. BMC Microbiol. 2024;24(1):467. doi:10.1186/s12866-024-03619-4
Clayton C, Kleanthous H, Coates P, Morgan D, Tabaqchali S. Sensitive detection of Helicobacter pylori by PCR. J Clin Microbiol. 1992;30(1):192-200. doi:10.1128/jcm.30.1.192-200.1992
Shallal ZS. Comparison between serological and molecular detection of Helicobacter pylori. Acad Int J Med Update. 2025;3(1):1-10. doi:10.59675/U311
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 European Journal of Clinical and Experimental Medicine
This work is licensed under a Creative Commons Attribution 4.0 International License.
Our open access policy is in accordance with the Budapest Open Access Initiative (BOAI) definition: this means that articles have free availability on the public Internet, permitting any users to read, download, copy, distribute, print, search, or link to the full texts of these articles, crawl them for indexing, pass them as data to software, or use them for any other lawful purpose, without financial, legal, or technical barriers other than those inseparable from having access to the Internet itself.
All articles are published with free open access under the CC-BY Creative Commons attribution license (the current version is CC-BY, version 4.0). If you submit your paper for publication by the Eur J Clin Exp Med, you agree to have the CC-BY license applied to your work. Under this Open Access license, you, as the author, agree that anyone may download and read the paper for free. In addition, the article may be reused and quoted provided that the original published version is cited. This facilitates freedom in re-use and also ensures that Eur J Clin Exp Med content can be mined without barriers for the research needs.
