OBJECTIVE: To explore the possibility that an analysis of antibody specificity to separated components of mycobacteria in a group of tuberculous patients may reveal a combination of target antigens whose antibodies could form the basis of a useful serodiagnostic test. DESIGN: Immunoblots of 1-dimensional (SDS-PAGE) and 2-dimensional (isoelectric focusing/SDS-PAGE) separation of antigenic extracts of Mycobacterium tuberculosis H37Rv (MTSE) and M. bovis bacille Calmette-Guérin (BCG) (MBSE) with 52 tuberculous and 59 BCG-vaccinated control human sera were analyzed for band and spot reactivity patterns that are indicative of infection with M. tuberculosis. RESULTS: Reactivity to antigens banding in the 10-18 kDa, 37-43 kDa and 70-90 kDa regions allowed a good discrimination between patients and normal subjects. Patients' sera reacting with antigens in the 22-30 and 70-88 kDa regions differentiated responses to MTSE and MBSE. In 2-D immunoblotting, patients' sera only reacted with antigens separating at approximately pI 6.5/26-28 kDa, pI 4.8/38 kDa and pI 6.5/70-79 kDa position and the responses were specific for M. tuberculosis (MTSE). CONCLUSION: These results provide evidence that a combination of these M. tuberculosis antigens may be a useful basis for developing a diagnostic antibody test. Additionally, they may help to define antigens, and host antibody responses that are specific to one but not the other of the two closely related species.
OBJECTIVE: To explore the possibility that an analysis of antibody specificity to separated components of mycobacteria in a group of tuberculouspatients may reveal a combination of target antigens whose antibodies could form the basis of a useful serodiagnostic test. DESIGN: Immunoblots of 1-dimensional (SDS-PAGE) and 2-dimensional (isoelectric focusing/SDS-PAGE) separation of antigenic extracts of Mycobacterium tuberculosis H37Rv (MTSE) and M. bovis bacille Calmette-Guérin (BCG) (MBSE) with 52 tuberculous and 59 BCG-vaccinated control human sera were analyzed for band and spot reactivity patterns that are indicative of infection with M. tuberculosis. RESULTS: Reactivity to antigens banding in the 10-18 kDa, 37-43 kDa and 70-90 kDa regions allowed a good discrimination between patients and normal subjects. Patients' sera reacting with antigens in the 22-30 and 70-88 kDa regions differentiated responses to MTSE and MBSE. In 2-D immunoblotting, patients' sera only reacted with antigens separating at approximately pI 6.5/26-28 kDa, pI 4.8/38 kDa and pI 6.5/70-79 kDa position and the responses were specific for M. tuberculosis (MTSE). CONCLUSION: These results provide evidence that a combination of these M. tuberculosis antigens may be a useful basis for developing a diagnostic antibody test. Additionally, they may help to define antigens, and host antibody responses that are specific to one but not the other of the two closely related species.