Astudy on mycobacterial proteins in the serodiagnosis of tuberculosis using ELISA and western blot |
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| Authors: | Ajay Kumar T A Venkita Subramanian Usha P Sharma |
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| Institution: | 1. Department of Biochemistry, Vallabhbhai Patel Chest Institute, University of Delhi, 110 007, Delhi
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| Abstract: | Protein fraction termed P2 was obtained from sonicates of Mycobacterium smegmatis, subjected to ammonium sulphate precipitation. P2 fraction was further fractionated into 4 fractions (PeakI-IV) by DEAE cellulose ion-exchange chromatography. ELISA was performed
on the sera of 104 tuberculous cases and 62 controls, using P2 fractions Peak I as test antigens. Though the mean ELISA values in tuberculous cases were higher than the controls, no statistically
significant difference was found between the two. Antigens P2, Peak I and IV were tested on Western blots with pooled sera from tuberculous cases as well as controls. P2 fraction, Peak-I and IV were separated on PAGE-SDS, electroblotted onto nitrocellulose Sheets and the blots were subjected
to ELISA. Peak IV appeared as a single band (M.W. 55,000). P2 fraction exhibited some discriminatory bands on development of blots. |
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