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Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis

Academic Article
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Overview

authors

  • Landowski, C. P.
  • Godfrey, H. P.
  • Bentley-Hibbert, S. I.
  • Liu, X. Y.
  • Huang, Z. S.
  • Sepulveda, R.
  • Huygen, K.
  • Gennaro, M. L.
  • Moy, F. H.
  • Lesley, Scott

publication date

  • July 2001

journal

  • Journal of Clinical Microbiology  Journal

abstract

  • Laboratory diagnosis of tuberculosis is often difficult. Immunodetection of circulating Mycobacterium tuberculosis proteins shed during active infection would not depend on an intact host immune response and could take advantage of the speed and low costs afforded by antibody-based assays. We previously showed that patients with active tuberculosis had increased levels of circulating antigen 85 (Ag85) proteins independent of their tuberculin skin test status (S. I. Bentley-Hibbert, X. Quan, T. Newman, K. Huygen, and H. P. Godfrey, Infect. Immun. 67:581-588, 1999). To extend these observations to a Mycobacterium bovis BCG-vaccinated population and to another secreted mycobacterial protein, Ag85 and PstS-1 (protein antigen B, p38 antigen) were quantified in sera from 97 Chilean tuberculosis patients and healthy controls (many of whom had received BCG as children) using dot immunobinding, mouse monoclonal anti-BCG Ag85 complex antibody, and chicken antipeptide antibodies reactive with M. tuberculosis Ag85B and PstS-1. The latter antibodies had been raised to peptide-derived immunogens expressed on a novel proprietary protein carrier in Escherichia coli. Median serum Ag85 levels measured by using either anti-Ag85 antibody were significantly higher in patients with active tuberculosis than in healthy controls (P, <0.001 to 0.01); the median serum PstS-1 levels were similar in patients and controls. The sensitivity of significantly elevated circulating Ag85 levels in patients with pulmonary tuberculosis measured by anti-Ag85 complex or anti-Ag85B antibodies was 60 and 55%, respectively, but increased to 77% when results obtained with both anti-Ag85 antibodies were considered jointly (P < 0.02). The corresponding specificities for individual and joint consideration were 95, 85, and 80%, respectively. These results indicate that elevated Ag85 levels can be detected in patients with active tuberculosis even after BCG vaccination and suggest that combinatorial use of antibodies directed at different epitopes of this protein could provide a viable strategy for developing new host immune response-independent diagnostic tests for tuberculosis.

subject areas

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Antibodies, Bacterial
  • Antigens, Bacterial
  • BCG Vaccine
  • Female
  • Humans
  • Immunoblotting
  • Male
  • Middle Aged
  • Mycobacterium tuberculosis
  • Sensitivity and Specificity
  • Tuberculosis, Pulmonary
  • Vaccination
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Identity

International Standard Serial Number (ISSN)

  • 0095-1137

Digital Object Identifier (DOI)

  • 10.1128/jcm.39.7.2418-2424.2001

PubMed ID

  • 11427548
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Additional Document Info

start page

  • 2418

end page

  • 2424

volume

  • 39

issue

  • 7

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