Combined use of Western blot/ELISA to improve the serological diagnosis of human tuberculosis

Abstract

Two recombinant antigens and a crude bacterial antigen of a wild M. tuberculosis strain were used to detect specific IgG antibodies in sera from 52 patients with pulmonary tuberculosis, confirmed by an acid-fast smear and serum culture of these patients and that of 25 contacts. The patients were not infected with HIV. We evaluated the sensitivity and specificity of ELISA, based on the recombinant TbF6® ® ® ® ® and TbF6/DPEP antigen and a search for reactivity patterns in the Western blot technique, using whole mycobacterium antigen. Serum samples from 22 healthy individuals and from 30 patients with lung diseases other than tuberculosis were used as controls. The best ELISA results were obtained with the TbF6/DPEP antigen combination, which gave 85% sensitivity and 91% specificity. ELISA sensitivity improved from 85% to 92% when the Western blot results were used. Western blot specificity was 100% when antibody reactivity with different antigenic bands was analyzed and associated. The association of TbF6/DPEP antigens used in ELISA with specific patterns of reactivity determined by Western blot can help make an identification when classic methods for the diagnosis of pulmonary tuberculosis are not sufficient.

Comparison of the Proteosomes and Antigenicities of Secreted and Cellular Proteins Produced byMycobacterium paratuberculosis

“…Beck et al found at least four serum antibody response profiles when testing human tuberculosis patient sera against M. tuberculosis antigens (2,3). Serum antibody responses are primarily affected by the stage of infection.…”

Assessment of Three Commercially Available Serologic Assays for Detection of Antibodies to Mycobacterium tuberculosis and Identification of Active Tuberculosis

“…These shortcomings result in the unnecessary treatment of many individuals who may not need it (3,17,32,45). While the tuberculin skin test (TST) and the QuantiFERON-TB Gold (QFT-G), the traditional methods for latent TB infection screening, rely on the cell-mediated response, the humoral response appears to correlate with the progression of the infection to active TB disease (5,6,11,15,20,21).Many studies have been conducted to evaluate the utility of individual specific Mycobacterium tuberculosis antigens for detecting antibodies in patients with active TB disease (1,7,10,11,20,21,25,26,29,38,39,45,46). Several of these antigens have been developed into commercial assays capable of detecting M. tuberculosis antibodies (4,28,35,53).…”

“…Many studies have been conducted to evaluate the utility of individual specific Mycobacterium tuberculosis antigens for detecting antibodies in patients with active TB disease (1,7,10,11,20,21,25,26,29,38,39,45,46). Several of these antigens have been developed into commercial assays capable of detecting M. tuberculosis antibodies (4,28,35,53).…”

Recombinant cold shock domain containing protein is a potential antigen to detect specific antibody during early and late infections of Haemonchus contortus in goat

“…It is further added that H. contortus interaction with host starts after the transition of L 3 into L 4 stage [28]. Furthermore, recombinant protein, having high immunoreactive concentration, may act as best immunodiagnostic antigen by achieving high sensitivity and specificity [29][30][31][32][33][34][35][36]. Recombinant Hc-CS is a major constituent of HcES proteins [23] which may have potential to diagnose H. contortus at early stage of infection.…”