Comparison of A60 and three glycolipid antigens in an ELISA test for tuberculosis.

OBJECTIVES

To compare the diagnostic usefulness in tuberculosis of the serodiagnostic enzyme-linked immunosorbent assay (ELISA) kit A60 (Anda Biologicals, Strasbourg, France) and of our domestic ELISA based on three purified cell wall glycolipid antigens.

METHODS

The presence and concentrations of IgG and IgM anti-A60 antibodies and anti-LOS, anti-DAT and anti-PGLTb1 antibodies against the glycolipid antigens were determined by ELISA in 50 HIV-seronegative and 46 HIV-seropositive patients, with documented active tuberculosis. The specificity of these ELISAs was determined with use of sera from 50 healthy blood donors, 29 patients with non-mycobacterial pulmonary diseases and 24 HIV-positive patients with disseminated Mycobacterium avium infection.

RESULTS

With a calculated cut-off for each antigen and immunoglobulin that gave a specificity higher than or equal to 98%, the cumulative ELISA results showed that only 36.5% of the patients with tuberculosis had a positive response in the A60 test, as compared with 84.4% who showed a response to the three glycolipid antigens (p<0.001). This striking difference persisted when the cumulative sensitivities were calculated according to the HIV status of the patients and the localization of tuberculosis. The anti-A60 antibody (IgG and IgM) levels and the degree of sensitivity of the ELISA for detection of A60 antigen were always lower in HIV-positive patients with pulmonary and extrapulmonary tuberculosis than in HIV-negative patients with tuberculosis. The sensitivity of A60 ELISA was further decreased in HIV-positive patients with low CD4+ lymphocytes counts, in contrast to the results with the three glycolipid antigens.

CONCLUSIONS

These results show the limitations of the A60 ELISA, and confirm the potencies of the glycolipid antigens in serodiagnosis of tuberculosis in HIV-positive and HIV-negative patients.