Isolation and evaluation of diagnostic value of two major secreted proteins of Mycobacterium tuberculosis.

Senthil Kumar, K.S.; Uma Devi, K.R.; Raja Alamelu.

Indian Journal of Chest Disease and Allied Sciences; 2002; 44; 225-232.

Two secreted antigens of Mycobacterium tuberculosis , namely the antigen 85 complex (30/31) and 38 kDa antigens, were purified from the whole culture filtrate by using two dimensional preparative electrophoresis and anion exchange chromatography, respectively. Individual components of the antigen 85 complex namely, antigen 85A, 85B and 85C, were separated using hydrophobic interaction chromatography. The humoral antibody activity to these antigens in sputum positive cases of active pulmonary tuberculosis and normal healthy volunteers was determined by enzyme linked immunosorbent assay (ELISA) and immunoblot. Recombinant 38 kDa and antigen 6 were used as reference antigens for the assay. None of the healthy volunteers reacted with the 38 kDa antigen, while 52% of the TB sera reacted with it. Of the three components of the antigen 85 complex, 85B gave the highest positivity of 40 per cent. The results of combination of 38 kDa with antigen 6 offered better results with 76% positivity.

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