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. 1976 Apr 1;143(4):741–758. doi: 10.1084/jem.143.4.741

The serological classification of Neisseria gonorrhoeae. I. Isolation of the outer membrane complex responsible for serotypic specificity

PMCID: PMC2190160  PMID: 56416

Abstract

Neisseria gonorrhoeae has been subdivided into several classes of serological distinct groups. The serotyping system is based upon the antigenic specificity of a protein serotype antigen. This protein is the major polypeptide of the outer membrane of the gonococcus and accounts for over 60% of that membrane's total protein. The serotype antigen complex was isolated by mild extraction of intact organisms in 200 mM lithium acetate buffer, pH 6.0 with 10 mM EDTA for 2 h at 45 degrees C. The extract was fractionated on Sepharose 6B and partially purified by precipitation at pH 4.2 by addition of 10% (vol/vol) acetic acid. Each serotype antigen has a unique subunit molecular size as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Preliminary typing of a gonococcal strain may be performed by comparative SDA-PAGE. To date, 16 different serotypes, representing a diverse distribution, have been isolated.

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Selected References

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  1. Apicella M. A., Allen J. C. Isolation and characterization of the antigen of Neisseria gonorrhoeae. Infect Immun. 1973 Mar;7(3):315–321. doi: 10.1128/iai.7.3.315-321.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bragg P. D., Hou C. Organization of proteins in the native and reformed outer membrane of Escherichia coli. Biochim Biophys Acta. 1972 Aug 9;274(2):478–488. doi: 10.1016/0005-2736(72)90193-9. [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. Buchanan T. M., Swanson J., Holmes K. K., Kraus S. J., Gotschlich E. C. Quantitative determination of antibody to gonococcal pili. Changes in antibody levels with gonococcal infection. J Clin Invest. 1973 Nov;52(11):2896–2909. doi: 10.1172/JCI107486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carifo K., Catlin B. W. Neisseria gonorrhoeae auxotyping: differentiation of clinical isolates based on growth responses on chemically defined media. Appl Microbiol. 1973 Sep;26(3):223–230. doi: 10.1128/am.26.3.223-230.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Casper W. A. The Serological Classification of Gonococci by Comparative Agglutination. J Bacteriol. 1937 Oct;34(4):353–379. doi: 10.1128/jb.34.4.353-379.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chacko C. W., Nair G. M. Sero-diagnosis of gonorrhoea with a microprecipitin test using a lipopolysaccharide antigen from N. gonorrhoeae. Br J Vener Dis. 1969 Mar;45(1):33–39. doi: 10.1136/sti.45.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DANIELSSON D. THE DEMONSTRATION OF N. GONORRHOEAE WITH THE AID OF FLUORESCENT ANTIBODIES. 4. STUDIES BY IMMUNOFLUORESCENCE AND DOUBLE DIFFUSION-IN-GEL TECHNIQUE ON THE ANTIGENIC RELATIONSHIP BETWEEN N. GONORRHOEAE AND OTHER NEISSERIA STRAINS. Acta Pathol Microbiol Scand. 1965;64:267–276. doi: 10.1111/apm.1965.64.2.267. [DOI] [PubMed] [Google Scholar]
  10. DEACON W. E., PEACOCK W. L., Jr, FREEMAN E. M., HARRIS A. Identification of Neisseria gonorrhoeae by means of fluorescent antibodies. Proc Soc Exp Biol Med. 1959 Jun;101(2):322–325. doi: 10.3181/00379727-101-24925. [DOI] [PubMed] [Google Scholar]
  11. Devoe I. W., Gilchrist J. E. Release of endotoxin in the form of cell wall blebs during in vitro growth of Neisseria meningitidis. J Exp Med. 1973 Nov 1;138(5):1156–1167. doi: 10.1084/jem.138.5.1156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Frasch C. E., Gotschlich E. C. An outer membrane protein of Neisseria meningitidis group B responsible for serotype specificity. J Exp Med. 1974 Jul 1;140(1):87–104. doi: 10.1084/jem.140.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. GREAVES R. I. Preservation of living cells by freeze-drying. Ann N Y Acad Sci. 1960 Apr 13;85:723–728. doi: 10.1111/j.1749-6632.1960.tb49992.x. [DOI] [PubMed] [Google Scholar]
  14. Glynn A. A., Ward M. E. Nature and Heterogeneity of the Antigens of Neisseria gonorrhoeae Involved in the Serum Bactericidal Reaction. Infect Immun. 1970 Aug;2(2):162–168. doi: 10.1128/iai.2.2.162-168.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hill J. C., Peterson N. R., Weiss E. Characterization of spheroplast membranes of Neisseria meningitidis group B. Infect Immun. 1972 Apr;5(4):612–621. doi: 10.1128/iai.5.4.612-621.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hutchinson R. I. Typing the gonococcus. Br Med J. 1970 Jul 11;3(5714):107–107. doi: 10.1136/bmj.3.5714.107-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Inouye M., Yee M. L. Homogeneity of envelope proteins of Escherichia coli separated by gel electrophoresis in sodium dodecyl sulfate. J Bacteriol. 1973 Jan;113(1):304–312. doi: 10.1128/jb.113.1.304-312.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jephcott A. E., Reyn A. Neisseria gonorrhoeae. Colony variation I. Acta Pathol Microbiol Scand B Microbiol Immunol. 1971;79(5):609–614. doi: 10.1111/j.1699-0463.1971.tb00088.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kellogg D. S., Jr, Cohen I. R., Norins L. C., Schroeter A. L., Reising G. Neisseria gonorrhoeae. II. Colonial variation and pathogenicity during 35 months in vitro. J Bacteriol. 1968 Sep;96(3):596–605. doi: 10.1128/jb.96.3.596-605.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kellogg D. S., Jr, Turner E. M. Rapid fermentation confirmation of Neisseria gonorrhoeae. Appl Microbiol. 1973 Apr;25(4):550–552. doi: 10.1128/am.25.4.550-552.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  24. La Scolea L. J., Jr, Young F. E. Development of a defined minimal medium for the growth of Neisseria gonorrhoeae. Appl Microbiol. 1974 Jul;28(1):70–76. doi: 10.1128/am.28.1.70-76.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maeland J. A., Kristoffersen T. Immunochemical investigations on Neisseria gonorrhoeae endotoxin. 1. Characterization of phenol-water extracted endotoxin and comparison with aqueous ether preparations. Acta Pathol Microbiol Scand B Microbiol Immunol. 1971;79(2):226–232. [PubMed] [Google Scholar]
  26. Miller C. P., Boor A. K. THE CARBOHYDRATES OF GONOCOCCUS AND MENINGOCOCCUS : I. THE ALCOHOL-PRECIPITABLE FRACTION. J Exp Med. 1934 Jan 1;59(1):75–82. doi: 10.1084/jem.59.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moldow C., Robertson J., Rothfield L. Purification of bacterial membrane proteins. The use of guanidinium thiocyanate and urea. J Membr Biol. 1972;10(2):137–152. doi: 10.1007/BF01867850. [DOI] [PubMed] [Google Scholar]
  28. Novotny P., Turner W. H. Immunological heterogeneity of pili of Neisseria gonorrhoeae. J Gen Microbiol. 1975 Jul;89(1):87–92. doi: 10.1099/00221287-89-1-87. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  31. REYN A. Serological studies on gonococci; thermostability and bio-chemical nature of the gonococcus antigens; durability of the factor sera. Acta Pathol Microbiol Scand. 1949;26(2):252–268. [PubMed] [Google Scholar]
  32. Schnaitman C. A. Comparison of the envelope protein compositions of several gram-negative bacteria. J Bacteriol. 1970 Dec;104(3):1404–1405. doi: 10.1128/jb.104.3.1404-1405.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schnaitman C. A. Outer membrane proteins of Escherichia coli. I. Effect of preparative conditions on the migration of protein in polyacrylamide gels. Arch Biochem Biophys. 1973 Aug;157(2):541–552. doi: 10.1016/0003-9861(73)90673-5. [DOI] [PubMed] [Google Scholar]
  34. Schnaitman C. A. Outer membrane proteins of Escherichia coli. II. Heterogeneity of major outer membrane polypeptides. Arch Biochem Biophys. 1973 Aug;157(2):553–560. doi: 10.1016/0003-9861(73)90674-7. [DOI] [PubMed] [Google Scholar]
  35. Stokinger H. E., Ackerman H., Carpenter C. M. Studies on the gonococcus: II. Properties of an antigenic fraction isolated from cell-free gonococcal broth supernatants. J Bacteriol. 1944 Feb;47(2):141–147. doi: 10.1128/jb.47.2.141-147.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stokinger H. E., Carpenter C. M., Plack J. Studies on the gonococcus: III. Quantitative agglutinative reactions of the Neisseria with special reference to Neisseria gonorrhoeae. J Bacteriol. 1944 Feb;47(2):149–157. doi: 10.1128/jb.47.2.149-157.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Swanson J., Kraus S. J., Gotschlich E. C. Studies on gonococcus infection. I. Pili and zones of adhesion: their relation to gonococcal growth patterns. J Exp Med. 1971 Oct 1;134(4):886–906. doi: 10.1084/jem.134.4.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. THAYER J. D., MARTIN J. E., Jr A SELECTIVE MEDIUM FOR THE CULTIVATION OF N. GONORRHOEAE AND N. MENINGITIDIS. Public Health Rep. 1964 Jan;79:49–57. [PMC free article] [PubMed] [Google Scholar]
  39. Tramont E. C., Sadoff J. C., Artenstein M. S. Cross-reactivity of Neisseria gonorrhoeae and Neisseria meningitidis and the nature of antigens involved in the bactericidal reaction. J Infect Dis. 1974 Sep;130(3):240–247. doi: 10.1093/infdis/130.3.240. [DOI] [PubMed] [Google Scholar]
  40. WILSON J. F. A serological study of Neisseria gonorrhoeae. J Pathol Bacteriol. 1954 Oct;68(2):495–510. doi: 10.1002/path.1700680222. [DOI] [PubMed] [Google Scholar]
  41. Walstad D. L., Reitz R. C., Sparling P. F. Growth inhibition among strains of Neisseria gonorrhoeae due to production of inhibitory free fatty acids and lysophosphatidylethanolamine: absence of bacteriocins. Infect Immun. 1974 Sep;10(3):481–488. doi: 10.1128/iai.10.3.481-488.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. Wolf-Watz H., Elmros T., Normark S., Bloom G. D. Cell envelope of Neisseria gonorrhoeae: outer membrane and peptidoglycan composition of penicillin-sensitive and-resistant strains. Infect Immun. 1975 Jun;11(6):1332–1341. doi: 10.1128/iai.11.6.1332-1341.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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