A NEW QUANTITATIVE DETECTION METHOD OF RECOMBINANT CFP10-ESAT6 AMALGAMATION PROTEINS FROM MYCOBACTERIUM TUBERCULOSIS BASED ON MICRO-MAGNETIC PROBES STRATEGY

摘要

A new rapid, specific and sensitive method for assay of recombinant CFP10-ESAT6 amalgamation proteins from Mycobacterium tuberculosis was proposed. The method used streptavidin-coated magnetic beads to enrich the specific biotinylated anti-CFP10 antibody, then adopted a sandwich-type enzyme linked immunosorbent assay technology with two kinds of monoclonal antibodies: biotinylated anti-CFP10 antibody and HRP-labeled anti-CFP10 antibody to identify the target CFP10-ESAT6 proteins, and finally detected chemiluminescence intensity by a small home-made optical sensor. It was shown that, the corresponding chemiluminescence intensity had a good logarithmic linear response to the concentration of CFP10-ESAT6 proteins when ranging at 1 ~ 1000 ng/mL, and the correlation coefficient is 0.9937. The proposed method could detect the CFP10-ESAT6 proteins with low detection limit (1 ng/mL) and the detection time could be controlled within 45 min. Compared with commonly used detection methods of M. tuberculosis, this met...