• Logo
  • SBMUJournals

Designing a novel ELISA method to increase sensitivity and specificity of Helicobacter pylori whole cell antigen detection

Farideh Kamarehei, Alireza Khabiri, Massoud Saidijam, Meysam Soleimani, Mohammad Yousef Alikhani
130

Views

PDF

Abstract

Aim: In this research, we designed a direct Enzyme Linked Immunoassay method to detect Helicobacter pylori antigens in stool
specimens.
Background: Helicobacter pylori infection as the worldwide problem is related to many gastrointestinal disorders such as gastritis,
gastric cancer, non-ulcer disease, peptic ulcer disease and duodenal ulcer.
Methods: We produced and purified recombinant CagA and NapA antigens in Escherichia coli and extracted their antibodies from a
panel of positive sera specimens. We designed a novel enzyme linked immunoassay direct method in combination with the whole cell
for the qualitative and quantitative detection of Helicobacter pylori antigens in human stool. Assay performance was evaluated by
histopathology staining and urease activity.
Results: The sensitivity and specificity of assay was determined as 91.7 [95% confidence interval: 89.3–95.6%] and 93.1% [95% CI:
91.2–96.4%], respectively. Novel ELISA exhibits enhanced sensitivity and specificity of Helicobacter pylori detection in comparison
with another commercially available kit.
Conclusion: Combination of the recombinant antigens and whole cell of Helicobacter pylori in immunoassay designing is a new
approach about early diagnosis, treatment and fallowing up of the Helicobacter pylori infected patients, especially in peptic cancer
cases.
Keywords: CagA protein, Helicobacter pylori, Neutrophil activating protein A, Enzyme-linked immunosorbent assay.
(Please cite as: Kamarehei F, Khabiri AR, Saeedi Jam M, Soleimani M, Alikhani MY. Designing a novel ELISA
method based on CagA, NapA recombinant antigens to increase sensitivity and specificity of Helicobacter pylori
whole cell antigen detection. Gastroenterol Hepatol Bed Bench 2018;11(4):333-342).


Keywords

Helicobacter pylori; Cytotoxin-associated gene A; Neutrophil activating protein A; Enzyme-linked immunosorbent assay

References

Hooi JK, Lai WY, Ng WK, Suen MM, Underwood FE, Tanyingoh D, et al. Global prevalence of Helicobacter pylori infection: systematic review and meta-analysis. Gastroenterology. 2017;153(2):420-9.

https://www.ncbi.nlm.nih.gov/pubmed/28456631

Wotherspoon A, Ortiz-Hidalgo C, Falzon M, Isaacson P. Helicobacter pylori-associated gastritis and primary B-cell gastric lymphoma. Lancet. 1991;338(8776):1175-6.

https://www.ncbi.nlm.nih.gov/pubmed/1682595

Hunt R, Xiao S, Megraud F, Leon-Barua R, Bazzoli F, Van Der Merwe S, et al. Helicobacter pylori in developing countries. World gastroenterology organisation global guideline. J Gastrointestin Liver Dis. 2011;20(3):299-304.

https://www.ncbi.nlm.nih.gov/pubmed/21961099

Brown LM. Helicobacter pylori: epidemiology and routes of transmission. Epidemiol rev. 2000;22(2):283-97.

https://www.ncbi.nlm.nih.gov/pubmed/11218379

Miftahussurur M, Yamaoka Y. Diagnostic methods of Helicobacter pylori infection for epidemiological studies: critical importance of indirect test validation. BioMed res int. 2016;2016.

https://www.hindawi.com/journals/bmri/2016/4819423/

Gisbert JP, De La Morena F, Abraira V. Accuracy of monoclonal stool antigen test for the diagnosis of H. pylori infection: a systematic review and meta-analysis. Am J Gastroenterol. 2006;101(8):1921-30.

https://www.ncbi.nlm.nih.gov/pubmed/16780557

Wilcox M, Dent T, Hunter J, Gray J, Brown D, Wight D, et al. Accuracy of serology for the diagnosis of Helicobacter pylori infection--a comparison of eight kits. J Clin Pathol. 1996;49(5):373-6.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC500474/

Mégraud F, Lehours P. Helicobacter pylori detection and antimicrobial susceptibility testing. Clin Microbiol Rev. 2007;20(2):280-322.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1865594/

Chey WD, Wong BC. American College of Gastroenterology guideline on the management of Helicobacter pylori infection. Am J Gastroenterol. 2007;102(8):1808-25.

https://www.ncbi.nlm.nih.gov/pubmed/17608775

Kodama M, Murakami K, Okimoto T, Fukuda Y, Shimoyama T, Okuda M, et al. Influence of proton pump inhibitor treatment on Helicobacter pylori stool antigen test. World J Gastroenterol. 2012;18(1):44-48.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3251804/

Jones KR, Joo YM, Jang S, Yoo Y-J, Lee HS, Chung I-S, et al. Polymorphism in the CagA EPIYA motif impacts development of gastric cancer. J Clin Microbiol. 2009;47(4):959-68.

http://jcm.asm.org/content/47/4/959.full

Stein M, Rappuoli R, Covacci A. Tyrosine phosphorylation of the Helicobacter pylori CagA antigen after cag-driven host cell translocation. Proc Natl Acad Sci. 2000;97(3):1263-8.

http://www.pnas.org/content/97/3/1263

Fu H-W. Helicobacter pylori neutrophil-activating protein: from molecular pathogenesis to clinical applications. World J Gastroenterol. 2014;20(18):5294-301.

https://www.ncbi.nlm.nih.gov/pubmed/24833859

Farjadi V, Abtahi H, Zolfaghari M, Soufian S, Hasanzadeh L. Production of recombinant CagA protein of Helicobacter pylori. J Arak Uni Med Sci. 2013;16(7):36-44.

http://amuj.arakmu.ac.ir/browse.php?a_id=2336&sid=1&slc_lang=en

Yee Y, Yip K, Que T, Chang K, Li K, Lee C, et al. Efficacy of enzyme immunoassay for the detection of Helicobacter pylori antigens in frozen stool specimens: local validation. Aliment Pharmacol Ther. 2002;16(10):1739-42.

https://www.ncbi.nlm.nih.gov/pubmed/12269966

Armbruster DA, Pry T. Limit of blank, limit of detection and limit of quantitation. Clin Biochem Rev. 2008;29(Suppl 1):S49-52.

https://www.ncbi.nlm.nih.gov/pubmed/18852857

Zhu L, He J, Cao X, Huang K, Luo Y, Xu W. Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food. Sci Rep. 2016;6:16092.

https://www.ncbi.nlm.nih.gov/pubmed/26976753

Tehrani ZR, Azadmanesh K, Mostafavi E, Gharibzadeh S, Soori S, Azizi M, et al. High avidity anti-integrase antibodies discriminate recent and non-recent HIV infection: Implications for HIV incidence assay. J Virol Methods. 2018;253:5-10.

https://www.ncbi.nlm.nih.gov/pubmed/29248529

Kim SS, Ruiz VE, Carroll JD, Moss SF. Helicobacter pylori in the pathogenesis of gastric cancer and gastric lymphoma. Cancer lett. 2011;305(2):228-38.

https://www.ncbi.nlm.nih.gov/pubmed/20692762

Gisbert JP, Pajares JM. Stool antigen test for the diagnosis of Helicobacter pylori infection: a systematic review. Helicobacter. 2004;9(4):347-68.

https://www.ncbi.nlm.nih.gov/pubmed/15270750

Shimoyama T, Sawaya M, Ishiguro A, Hanabata N, Yoshimura T, Fukuda S. Applicability of a rapid stool antigen test, using monoclonal antibody to catalase, for the management of Helicobacter pylori infection. J Gastroenterol. 2011;46(4):487-91.

https://www.ncbi.nlm.nih.gov/pubmed/21264478

Krogfelt KA, Lehours P, Mégraud F. Diagnosis of Helicobacter pylori infection. Helicobacter. 2005;10:5-13.

https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1523-5378.2005.00341.x

Korkmaz H, Kesli R, Karabagli P, Terzi Y. Comparison of the diagnostic accuracy of five different stool antigen tests for the diagnosis of Helicobacter pylori infection. Helicobacter. 2013;18(5):384-91.

https://www.ncbi.nlm.nih.gov/pubmed/23551920

Veijola L, Oksanen A, Löfgren T, Sipponen P, Karvonen A-L, Rautelin H. Comparison of three stool antigen tests in confirming Helicobacter pylori eradication in adults. Scand j Gastroenterol. 2005;40(4):395-401.

https://www.ncbi.nlm.nih.gov/pubmed/16028433

Garza-González E, Perez-Perez GI, Maldonado-Garza HJ, Bosques-Padilla FJ. A review of Helicobacter pylori diagnosis, treatment, and methods to detect eradication. World j Gastroenterol. 2014;20(6):1438.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3925853/

Leal YA, Cedillo-Rivera R, Simón JA, Velázquez JR, Flores LL, Torres J. Utility of Stool Sample–based Tests for the Diagnosis of Helicobacter pylori Infection in Children. J Ped Gastroenterol Nut. 2011;52(6):718-28.

https://www.ncbi.nlm.nih.gov/pubmed/21478757

Kato S, Ozawa K, Okuda M, Fujisawa T, Kagimoto S, Konno M, et al. Accuracy of the stool antigen test for the diagnosis of childhood Helicobacter pylori infection: a multicenter Japanese study. Am J Gastroenterol. 2003;98(2):296-300.

https://www.ncbi.nlm.nih.gov/pubmed/12591044

Ni Y-H, Lin J-T, Huang S-F, Yang J-C, Chang M-H. Accurate diagnosis Helicobacter pylori infection by stool antigen test and 6 other currently available tests in children. J Pediatr. 2000;136(6):823-7.

https://www.ncbi.nlm.nih.gov/pubmed/10839883

Guo Y, Zhang S, Peng X, Zhan S. A systematic review of diagnosis of Helicobacter pylori infection by Helicobacter pylori stool antigen test. Chinese. 2005. 15;85(22):1564-7.

https://www.ncbi.nlm.nih.gov/pubmed/16179120

Kamarehei F, Ghaemi EA, Dadgar T. Prevalence of enterotoxin A and B genes in Staphylococcus aureus isolated from clinical samples and healthy carriers in Gorgan City, North of Iran. Indian J Pathol Microbiol. 2013;56(3):265-8.

https://www.ncbi.nlm.nih.gov/pubmed/24152506

Naseri Z, Alikhani MY, Hashemi SH, Kamarehei F, Arabestani MR. Prevalence of the most common virulence-associated genes among Brucella Melitensis isolates from human blood cultures in Hamadan Province, West of Iran. Iran J Med Sci. 2016;41(5):422-9.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4967487/

Kamarehei F, Rahimi-Alang S, Vaez H, Ghaemi E. Prevalence of Panton-valentine gene in Staphylococcus aureus isolated from clinical samples and healthy carriers in Gorgan city, north of Iran. Minerva biotecnol. 2015;27(1):51-4.

http://eprints.goums.ac.ir/4692/

Naseri Z, Kamarehei F, Hashemi SH, Moghimbeigi A, Eslami M, Asadi FT, et al. The seroprevalence of Brucellosis among blood donors in Hamadan province, west of Iran. J Pure Appl Microbiol. 2015;9(4):2865-70.

http://bloodjournal.ir/browse.php?a_id=745&sid=1&slc_lang=en

Dadgar T, Ghaemi E, Bahador N, Imani Fooladi A, Kamareie F. Detection of Staphylococcus Aureus Enterotoxin Genes AE. Med Lab J. 2014;7(5):1-8.

http://www.ijpmonline.org/article.asp?issn=0377-4929;year=2013;volume=56;issue=3;spage=265;epage=268;aulast=Kamarehei

Matsuo Y, Kido Y, Akada J, Shiota S, Binh TT, Trang TTH, et al. Novel CagA ELISA exhibits enhanced sensitivity of Helicobacter pylori CagA antibody. World J Gastroenterol. 2017;23(1):48.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5221286/




DOI: https://doi.org/10.22037/ghfbb.v11i4.1451