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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1986 Jul;24(1):126–130. doi: 10.1128/jcm.24.1.126-130.1986

Identification of Mycobacterium tuberculosis antigens in Seibert fractions by immunoblotting.

S R Coates, D Hansen, G Schecter, G Slutkin, P Hopewell, L Affronti, D F Echenberg
PMCID: PMC268846  PMID: 3088029

Abstract

Seibert fractions prepared from Mycobacterium tuberculosis culture filtrates were evaluated by immunoblotting with a serum pool from patients with active pulmonary tuberculosis. Antibody activity was observed primarily with antigens in the polysaccharide II and A protein fractions; these fractions were further evaluated by immunoblotting with sera from individual patients with tuberculosis, from individuals without tuberculosis and positive for the purified protein derivative antigen skin test, and from individuals negative for the purified protein derivative antigen skin test. The antigens identified in the protein A fraction, a 32,000-molecular-weight antigen and a heterogeneous high-molecular-weight antigen, reacted with antibody found in sera from all patients with tuberculosis and with antibody from over 25% of the control individuals. A 10,000-molecular-weight antigen, a 30,000- to 44,000-molecular-weight antigen, and a heterogeneous high-molecular-weight antigen were observed in the polysaccharide II fraction; these antigens reacted with serum antibody from 70% or more of the patients with tuberculosis and with antibody from 20 to 70% of the control individuals. One of the antigens, with a molecular weight ranging from 17,000 to 28,000 in the polysaccharide II fraction, reacted with antibody in 64% of the sera from patients with tuberculosis but with only 1 of 15 control normal sera. This antigen may elicit an antibody response specifically associated with tuberculosis.

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Selected References

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  1. Affronti L. F., Fife E. H., Grow L. Serodiagnostic test for tuberculosis. Am Rev Respir Dis. 1973 May;107(5):822–825. doi: 10.1164/arrd.1973.107.5.822. [DOI] [PubMed] [Google Scholar]
  2. Bardana E. J., Jr, McClatchy J. K., Farr R. S., Minden P. Universal occurrence of antibodies to tubercle bacilli in sera from non-tuberculous and tuberculous individuals. Clin Exp Immunol. 1973 Jan;13(1):65–77. [PMC free article] [PubMed] [Google Scholar]
  3. Benjamin R. G., Daniel T. M. Serodiagnosis of tuberculosis using the enzyme-linked immunoabsorbent assay (ELISA) of antibody to Mycobacterium tuberculosis antigen 5. Am Rev Respir Dis. 1982 Dec;126(6):1013–1016. doi: 10.1164/arrd.1982.126.6.1013. [DOI] [PubMed] [Google Scholar]
  4. Chakrabarty A. K., Maire M. A., Lambert P. H. SDS-PAGE analysis of M. leprae protein antigens reacting with antibodies from sera from lepromatous patients and infected armadillos. Clin Exp Immunol. 1982 Sep;49(3):523–531. [PMC free article] [PubMed] [Google Scholar]
  5. Chaparas S. D., Janicki B. W., Good R., Johnson A. H., Wright G., Goldstein R., Daniel T. M., Alling D. W. In vitro and in vivo reactivity and specificity of fractions from sonicates of Mycobacterium tuberculosis separated by gradient acrylamide gel electrophoresis. Am Rev Respir Dis. 1980 Oct;122(4):533–542. doi: 10.1164/arrd.1980.122.4.533. [DOI] [PubMed] [Google Scholar]
  6. Cole R. V., Lazarus A. W., Hedrick H. G. Development and evaluation of a simple latex agglutination test for diagnosis of tuberculosis. Appl Microbiol. 1972 Oct;24(4):525–534. doi: 10.1128/am.24.4.525-534.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Daniels T. M., Affronti L. F. Immunoelectrophoretic analysis of antigenic constituents of Seibert fractions of mycobacterial culture filtrates. Identification with the proposed United States-Japan reference nomenclature. Am Rev Respir Dis. 1973 Nov;108(5):1244–1248. doi: 10.1164/arrd.1973.108.5.1244. [DOI] [PubMed] [Google Scholar]
  8. Erlich H. A., Rodgers G., Vaillancourt P., Araujo F. G., Remington J. S. Identification of an antigen-specific immunoglobulin M antibody associated with acute Toxoplasma infection. Infect Immun. 1983 Aug;41(2):683–690. doi: 10.1128/iai.41.2.683-690.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hsu S. M., Soban E. Color modification of diaminobenzidine (DAB) precipitation by metallic ions and its application for double immunohistochemistry. J Histochem Cytochem. 1982 Oct;30(10):1079–1082. doi: 10.1177/30.10.6182185. [DOI] [PubMed] [Google Scholar]
  10. Kalish S. B., Radin R. C., Phair J. P., Levitz D., Zeiss C. R., Metzger E. Use of an enzyme-linked immunosorbent assay technique in the differential diagnosis of active pulmonary tuberculosis in humans. J Infect Dis. 1983 Mar;147(3):523–530. doi: 10.1093/infdis/147.3.523. [DOI] [PubMed] [Google Scholar]
  11. Minden P., McClatchy J. K., Cooper R., Bardana E. J., Jr, Farr R. S. Shared antigens between Mycobacterium bovis (BCG) and other bacterial species. Science. 1972 Apr 7;176(4030):57–58. doi: 10.1126/science.176.4030.57. [DOI] [PubMed] [Google Scholar]
  12. Mitchison D. A., Aber V. R., Ahmad F. J., Allen B. W., Devi S. Evaluation of a serological test for tuberculosis. Br Med J. 1977 May 28;1(6073):1383–1384. doi: 10.1136/bmj.1.6073.1383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Reggiardo Z., Vazquez E., Schnaper L. ELISA tests for antibodies against mycobacterial glycolipids. J Immunol Methods. 1980;34(1):55–60. doi: 10.1016/0022-1759(80)90224-0. [DOI] [PubMed] [Google Scholar]
  14. Stroebel A. B., Daniel T. M., Lau J. H., Leong J. C., Richardson H. Serologic diagnosis of bone and joint tuberculosis by an enzyme-linked immunosorbent assay. J Infect Dis. 1982 Aug;146(2):280–283. doi: 10.1093/infdis/146.2.280. [DOI] [PubMed] [Google Scholar]
  15. Toussaint A. J., Fife E. H., Parlett R. C., Affronti L. F., Wright G. L., Reich M., Morse W. C. A soluble antigen fluorescent antibody test for the serodiagnosis of Mycobacterium tuberculosis infection. Am J Clin Pathol. 1969 Dec;52(6):708–713. doi: 10.1093/ajcp/52.6.708. [DOI] [PubMed] [Google Scholar]
  16. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  17. Winters W. D., Cox R. A. Serodiagnosis of tuberculosis by radioimmunoassay. Am Rev Respir Dis. 1981 Nov;124(5):582–585. doi: 10.1164/arrd.1981.124.5.582. [DOI] [PubMed] [Google Scholar]
  18. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]
  19. Zeiss C. R., Radin R. C., Williams J. E., Levitz D., Phair J. P. Detection of immunoglobulin G antibody to purified protein derivative in patients with tuberculosis by radioimmunoassay and enzyme-linked immunosorbent assay. J Clin Microbiol. 1982 Jan;15(1):93–96. doi: 10.1128/jcm.15.1.93-96.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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