Abstract
We report the development of an EIA specific for antituberculosis antibody in human serum for the clinical evaluation of tuberculosis. We developed a TLC immunostaining method to detect specific antigens for antibodies in the serum of patients with tuberculosis. The detected specific antigens, TDM and specific glycolipid fraction, were individually purified from M. tuberculosis H37Rv by column chromatography. The two purified fractions were mixed and the mixture, termed TBGL antigen, was applied to an enzyme immunoassay suitable for the measurement of antituberculosis antibodies in serum. This EIA meets all the requirements of routine clinical assay in terms of sensitivity (detection limit: 0.125 U/ml), reproducibility (total CV: 3.3–6.0%), accuracy (recovery: 96–105%), simplicity and rapidity (<2.5 h). Clinical validation of the assay was confirmed by the measurement of the anti‐tuberculosis antibody in the serum of normal subjects and patients with pulmonary tuberculosis. The EIA tested in this study showed a high serodiagnostic discriminating power (90% sensitivity and 98% specificity). J. Clin. Lab. Anal. 11:140–145, 1997. © 1997 Wiley‐Liss, Inc.
Keywords: M. tuberculosis, pulmonary tuberculosis, serodiagnosis, cord factor, ELISA
References
- 1. Bloom BR, Murray CJL: Tuberculosis: Commentary on a reemergent killer. Science 257: 1055–1064, 1992. [DOI] [PubMed] [Google Scholar]
- 2. Hua H, Oka S, Yamamura Y, Kusunose E, Kusunose M, Yano I: Rapid serodiagnosis of human mycobacteriosis by ELISA using cord factor(trehalose‐6,6′‐dimycolate) purified from mycobacterium tuberculosis as antigen. FEMS Microbiol Lett 76: 201–204, 1991. [DOI] [PubMed] [Google Scholar]
- 3. Ishikawa E, Imagawa M, Hashida S, Yoshitake S, Hamaguchi Y, Ueno T: Enzyme‐labeling of antibodies and their fragments for enzyme immunoassay and immunohistochemical staining. J Immunoassay 4: 209–327, 1983. [DOI] [PubMed] [Google Scholar]
- 4. Kohno H, Sato S, Chiba H, Kobayashi K, Ikegawa S, Kurosawa T, Tohma M: Monoclonal antibodies specific for 18‐hydroxycortisol and their use in an enzyme immunoassay for human urinary 18‐hydrocortisol for diagnosis of primary aldosteronism. Clin Biochem 27: 227–282, 1994. [DOI] [PubMed] [Google Scholar]
- 5. Daniel TM, BeBanne SM: The serodiagnosis of tuberculosis and other mycobacterial diseases by enzyme‐linked immunosorbent assay. Am Rev Respir Dis 135: 1137–1151, 1987. [DOI] [PubMed] [Google Scholar]
- 6. Daniel TM: Antibody and antigen detection for the immunodiagnosis of tuberculosis: Why not? What more is needed? Where do we stand today. J Infect Dis 158: 679–680, 1988. [DOI] [PubMed] [Google Scholar]
- 7. Takahashi Y: Specific serum agglutination of kaolin particles sensitized with tubercle phosphatide and its clinical evaluation as a serodiagnostic test for tuberculosis. Am Rev Respir Dis 85: 708–719, 1962. [DOI] [PubMed] [Google Scholar]
- 8. Benjamin RG, Daniel TM: Serodiagnosis of tuberculosis using the enzyme‐linked immunosorbent assay (ELISA) of antibody to Mycobacterium tuberculosis antigen 5. Am Rev Respir Dis 126: 1023, 1982. [DOI] [PubMed] [Google Scholar]
- 9. Kusano N: Evaluation of serodiagnosis detecting IgG antibodies against purified protein derivative and alpha antigen in patients with active pulmonary tuberculosis by enzyme‐linked immunosorbent assay. Kekkaku 82: 211, 1987. [PubMed] [Google Scholar]
- 10. Kato M: Immunochemical properties of anti‐cord factor antibody: Effect of anti‐cord factor antibody on experimental tuberculosis in mice. Infect Immun 7: 9–12, 1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. Kato M: Antibody formation to trehalose‐6,6′‐dimycolate (cord factor) of mycobacterium tuberculosis. Infect Immun 5: 203–212, 1972. [DOI] [PMC free article] [PubMed] [Google Scholar]