Astudy on mycobacterial proteins in the serodiagnosis of tuberculosis using ELISA and western blot

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.