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Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1995 Mar;2(2):156–159. doi: 10.1128/cdli.2.2.156-159.1995

Isolation and preliminary evaluation of a low-molecular-mass antigen preparation for improved detection of Helicobacter pylori immunoglobulin G antibodies.

L P Andersen 1, F Espersen 1, A Souckova 1, M Sedlackova 1, A Soucek 1
PMCID: PMC170119  PMID: 7697523

Abstract

Previously, immunoglobulin G (IgG) antibodies to five antigens with a relative molecular mass of between 15 and 30 kDa from Helicobacter pylori were found to be significantly more frequent in H. pylori-infected patients than in noninfected patients. In this study, these specific low-molecular-mass (LMW) antigens were separated by ultrafiltration of whole-cell sonicates. The LMW antigen preparation was evaluated by enzyme-linked immunosorbent assay with serum samples from 76 children with abdominal symptoms and 151 adults with dyspeptic symptoms. H. pylori was cultured or seen in 40 (53%) children and 83 (55%) adults. Increased antibody levels to H. pylori were found in serum from 35 (46%) children and 88 (58%) adults. Values for sensitivity, specificity, and predictive value of positive and negative results of the test were higher with LMW antigens than with the heat-stable antigen previously described. The low specificity and predictive value of a positive result were due to seropositive results for 21 persons with a negative culture for H. pylori and negative microscopy results for Helicobacter-like organisms in biopsies from gastric mucosa. Histologically, chronic gastritis was demonstrated in 43% of these persons, and 19% had peptic ulcer, indicating that they have or have had H. pylori infection. Specific antibodies to H. pylori were confirmed in all 21 patients by the Western immunoblot technique. Use of the LMW antigen improved the IgG antibody detection in patients with H. pylori infection, even though the results reflect the difficulties in establishing a true gold standard for diagnosis of H. pylori infection.

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Selected References

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  1. Andersen L. P., Blom J., Nielsen H. Survival and ultrastructural changes of Helicobacter pylori after phagocytosis by human polymorphonuclear leukocytes and monocytes. APMIS. 1993 Jan;101(1):61–72. [PubMed] [Google Scholar]
  2. Andersen L. P., Elsborg L., Justesen T. Campylobacter pylori in peptic ulcer disease. III. Symptoms and paraclinical and epidemiologic findings. Scand J Gastroenterol. 1988 Apr;23(3):347–350. doi: 10.3109/00365528809093877. [DOI] [PubMed] [Google Scholar]
  3. Andersen L. P., Espersen F. Immunoglobulin G antibodies to Helicobacter pylori in patients with dyspeptic symptoms investigated by the western immunoblot technique. J Clin Microbiol. 1992 Jul;30(7):1743–1751. doi: 10.1128/jcm.30.7.1743-1751.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andersen L. P., Nielsen H. Peptic ulcer: an infectious disease? Ann Med. 1993 Dec;25(6):563–568. [PubMed] [Google Scholar]
  5. Andersen L. P., Raskov H., Elsborg L., Holck S., Justesen T., Fischer Hansen B., Møller Nielsen C., Gaarslev K. Prevalence of antibodies against heat-stable antigens from Helicobacter pylori in patients with dyspeptic symptoms and normal persons. APMIS. 1992 Sep;100(9):779–789. [PubMed] [Google Scholar]
  6. Biasco G., Paganelli G. M., Santucci R., Miglioli M., Barbara L. Cell proliferation kinetics as a marker of gastric cancer risk. Ital J Gastroenterol. 1991 Jan;23(1):24–28. [PubMed] [Google Scholar]
  7. Buchvald D., Maeland J. A. Characterization of a 25,000-dalton Helicobacter pylori protein, cross-reacting with a Campylobacter jejuni protein. APMIS. 1992 May;100(5):470–478. [PubMed] [Google Scholar]
  8. Czinn S., Carr H., Sheffler L., Aronoff S. Serum IgG antibody to the outer membrane proteins of Campylobacter pylori in children with gastroduodenal disease. J Infect Dis. 1989 Mar;159(3):586–589. doi: 10.1093/infdis/159.3.586. [DOI] [PubMed] [Google Scholar]
  9. Forman D. Helicobacter pylori infection: a novel risk factor in the etiology of gastric cancer. J Natl Cancer Inst. 1991 Dec 4;83(23):1702–1703. doi: 10.1093/jnci/83.23.1702. [DOI] [PubMed] [Google Scholar]
  10. Jensen A. K., Andersen L. P., Gaarslev K., Wachmann C. H. Comparison of four second generation kits for detection of IgG antibodies against Helicobacter pylori in adults. Zentralbl Bakteriol. 1993 Sep;280(1-2):221–226. doi: 10.1016/s0934-8840(11)80959-x. [DOI] [PubMed] [Google Scholar]
  11. Jensen A. K., Andersen L. P., Wachmann C. H. Evaluation of eight commercial kits for Helicobacter pylori IgG antibody detection. APMIS. 1993 Oct;101(10):795–801. [PubMed] [Google Scholar]
  12. Johansen H. K., Nørgaard A., Andersen L. P., Jensen P., Nielsen H., Høiby N. Cross-reactive antigens shared by Pseudomonas aeruginosa, Helicobacter pylori, Campylobacter jejuni, and Haemophilus influenzae may cause false-positive titers of antibody to H. pylori. Clin Diagn Lab Immunol. 1995 Mar;2(2):149–155. doi: 10.1128/cdli.2.2.149-155.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kosunen T. U., Seppälä K., Sarna S., Sipponen P. Diagnostic value of decreasing IgG, IgA, and IgM antibody titres after eradication of Helicobacter pylori. Lancet. 1992 Apr 11;339(8798):893–895. doi: 10.1016/0140-6736(92)90929-w. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Maaroos H. I., Kekki M., Sipponen P., Salupere V., Villako K. Grade of Helicobacter pylori colonisation, chronic gastritis and relative risks of contracting high gastric ulcers: a seven-year follow-up. Scand J Gastroenterol Suppl. 1991;186:65–72. doi: 10.3109/00365529109103989. [DOI] [PubMed] [Google Scholar]
  16. Mendall M. A., Goggin P. M., Molineaux N., Levy J., Toosy T., Strachan D., Northfield T. C. Childhood living conditions and Helicobacter pylori seropositivity in adult life. Lancet. 1992 Apr 11;339(8798):896–897. doi: 10.1016/0140-6736(92)90931-r. [DOI] [PubMed] [Google Scholar]
  17. Misiewicz J. J. The Sydney System: a new classification of gastritis. Introduction. J Gastroenterol Hepatol. 1991 May-Jun;6(3):207–208. doi: 10.1111/j.1440-1746.1991.tb01467.x. [DOI] [PubMed] [Google Scholar]
  18. Mégraud F., Lamouliatte H. Helicobacter pylori and duodenal ulcer. Evidence suggesting causation. Dig Dis Sci. 1992 May;37(5):769–772. doi: 10.1007/BF01296437. [DOI] [PubMed] [Google Scholar]
  19. Newell D. G. Identification of the outer membrane proteins of Campylobacter pyloridis and antigenic cross-reactivity between C. pyloridis and C. jejuni. J Gen Microbiol. 1987 Jan;133(1):163–170. doi: 10.1099/00221287-133-1-163. [DOI] [PubMed] [Google Scholar]
  20. Perez-Perez G. I., Dworkin B. M., Chodos J. E., Blaser M. J. Campylobacter pylori antibodies in humans. Ann Intern Med. 1988 Jul 1;109(1):11–17. doi: 10.7326/0003-4819-109-1-11. [DOI] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vandenplas Y., Blecker U., Devreker T., Keppens E., Nijs J., Cadranel S., Pipeleers-Marichal M., Goossens A., Lauwers S. Contribution of the 13C-urea breath test to the detection of Helicobacter pylori gastritis in children. Pediatrics. 1992 Oct;90(4):608–611. [PubMed] [Google Scholar]
  23. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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