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. 1980 Apr;28(1):101–110. doi: 10.1128/iai.28.1.101-110.1980

Antigenic diversity of the serotype antigen complex of Neisseria gonorrhoeae: analysis by an indirect enzyme-linked immunoassay.

K H Johnston
PMCID: PMC550898  PMID: 6769815

Abstract

An indirect enzyme-linked immunoassay (ELISA) has been developed to analyze the antigenic profile of the outer membrane serotype complex of Neisseria gonorrhoeae. Antisera raised in rabbits to serotype-specific vesicles (SSV) reacted primarily with homologous SSV; however, there was significant cross-reactivity (less than 50%) with heterologous SSV. N. meningitidis SSV cross-reacted with all antigonococcal SSV but at a lower degree (less than 20%). Preimmune sera did not cross-react significantly with all antigonoccoccal SSV. The sensitivity of the ELISA was enhanced when the integral SSV proteins 1a and 2 were used as adsorbed antigen. Heterologous anti-SSV cross-reacted slightly, having ELISA values less than 15% of the homologous reaction. Antisera prepared by immunoabsorbent affinity columns were highly specific. Homologous affinity anti-SSV reacted only with proteins 1a and 2. The reaction of immune sera was inhibited by homologous proteins 1a and 2; lipopolysaccharide and proteins 1a and 2 isolated from heterologous serotypes did not inhibit the reaction. The reaction of affinity-purified antisera could be inhibited only by homologous protein 1a. By the use of affinity-purified antisera, a specific and highly sensitive ELISA was developed to analyze the antigenic profile of strains of N. gonorrhoeae.

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Selected References

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  1. Apicella M. A. Antigenically distinct populations of Neisseria gonorrhoeae: isolation and characterization of the responsible determinants. J Infect Dis. 1974 Dec;130(6):619–625. doi: 10.1093/infdis/130.6.619. [DOI] [PubMed] [Google Scholar]
  2. Apicella M. A. Serogrouping of Neisseria gonorrhoeae: identification of four immunologically distinct acidic polysaccharides. J Infect Dis. 1976 Oct;134(4):377–383. doi: 10.1093/infdis/134.4.377. [DOI] [PubMed] [Google Scholar]
  3. Brodeur B. R., Ashton F. E., Diena B. B. Enzyme-linked immunosorbent assays for the detection of Neisseria gonorrhoeae specific antibodies. Can J Microbiol. 1978 Nov;24(11):1300–1305. doi: 10.1139/m78-211. [DOI] [PubMed] [Google Scholar]
  4. Buchanan T. M. Antigenic heterogeneity of gonococcal pili. J Exp Med. 1975 Jun 1;141(6):1470–1475. doi: 10.1084/jem.141.6.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carlsson H. E., Lindberg A. A., Hammarström S., Ljunggren A. Quantitation of Salmonella O-antibodies in human sera by enzyme-linked immunosorbent assay (ELISA). Int Arch Allergy Appl Immunol. 1975;48(4):485–494. doi: 10.1159/000231336. [DOI] [PubMed] [Google Scholar]
  6. Carlsson H. E., Lindberg A. A., Hammarström S. Titration of antibodies to salmonella O antigens by enzyme-linked immunosorbent assay. Infect Immun. 1972 Nov;6(5):703–708. doi: 10.1128/iai.6.5.703-708.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  8. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garten W., Henning U. Cell envelope and shape of Escherichia coli K12. Isolation and preliminary characterization of the major ghost-membrane proteins. Eur J Biochem. 1974 Sep 1;47(2):343–352. doi: 10.1111/j.1432-1033.1974.tb03699.x. [DOI] [PubMed] [Google Scholar]
  10. Glynn A. A., Ison C. Serological diagnosis of gonorrhoea by an enzyme-linked immunosorbent assay (Elisa). Br J Vener Dis. 1978 Apr;54(2):97–102. doi: 10.1136/sti.54.2.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heckels J. E. The surface of Neisseria gonorrhoeae: isolation of the major components of the outer membrane. J Gen Microbiol. 1977 Apr;99(2):333–341. doi: 10.1099/00221287-99-2-333. [DOI] [PubMed] [Google Scholar]
  12. Ichihara S., Mizushima S. Characterization of major outer membrane proteins O-8 and O-9 of Escherichia coli K-12. Evidence that structural genes for the two proteins are different. J Biochem. 1978 Apr;83(4):1095–1100. doi: 10.1093/oxfordjournals.jbchem.a131998. [DOI] [PubMed] [Google Scholar]
  13. Jodal U., Ahlstedt S., Carlsson B., Hanson L. A., Lindberg U., Sohl A. Local antibodies in childhood urinary tract infection: a preliminary study. Int Arch Allergy Appl Immunol. 1974;47(4):537–546. doi: 10.1159/000231248. [DOI] [PubMed] [Google Scholar]
  14. Johnston K. H., Gotschlich E. C. Isolation and characterization of the outer membrane of Neisseria gonorrhoeae. J Bacteriol. 1974 Jul;119(1):250–257. doi: 10.1128/jb.119.1.250-257.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  16. Nakamura K., Mizushima S. Effects of heating in dodecyl sulfate solution on the conformation and electrophoretic mobility of isolated major outer membrane proteins from Escherichia coli K-12. J Biochem. 1976 Dec;80(6):1411–1422. doi: 10.1093/oxfordjournals.jbchem.a131414. [DOI] [PubMed] [Google Scholar]
  17. OSBORN M. J. STUDIES ON THE GRAM-NEGATIVE CELL WALL. I. EVIDENCE FOR THE ROLE OF 2-KETO- 3-DEOXYOCTONATE IN THE LIPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM. Proc Natl Acad Sci U S A. 1963 Sep;50:499–506. doi: 10.1073/pnas.50.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Perry M. B., Daoust V. The lipopolysaccharides of Neisseria gonorrhoeae colony types 1 and 4. Can J Biochem. 1975 May;53(5):623–629. doi: 10.1139/o75-084. [DOI] [PubMed] [Google Scholar]
  19. Rosenbusch J. P. Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. J Biol Chem. 1974 Dec 25;249(24):8019–8029. [PubMed] [Google Scholar]
  20. Van Alphen L., Havekes L., Lugtenberg B. Major outer membrane protein d of Escherichia coli K12. Purification and in vitro activity of bacteriophages k3 and f-pilus mediated conjugation. FEBS Lett. 1977 Mar 15;75(1):285–290. doi: 10.1016/0014-5793(77)80104-x. [DOI] [PubMed] [Google Scholar]
  21. Van Weemen B. K., Schuurs A. H.W.M. Immunoassay using antigen-enzyme conjugates. FEBS Lett. 1971 Jun 24;15(3):232–236. doi: 10.1016/0014-5793(71)80319-8. [DOI] [PubMed] [Google Scholar]
  22. Veldkamp J., Visser A. M. Application of the enzyme-linked immunosorbent assay (ELISA) in the serodiagnosis of syphilis. Br J Vener Dis. 1975 Aug;51(4):227–231. doi: 10.1136/sti.51.4.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Voller A., Bidwell D. E., Bartlett A. Enzyme immunoassays in diagnostic medicine. Theory and practice. Bull World Health Organ. 1976;53(1):55–65. [PMC free article] [PubMed] [Google Scholar]
  24. 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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