Colorimetric Detection of MPT64 Antibody Based on an Aptamer Adsorbed Magnetic Nanoparticles for Diagnosis of Tuberculosis

Abstract

We have developed a colorimetric biosensing system for the detection of antibody against MPT64, a protein secreted by Mycobacterium tuberculosis, using aptamer DNA adsorbed Fe3O4 magnetic nanoparticles (MNPs) for diagnosis of tuberculosis (TB). In this system, MNPs were first incubated with single stranded (ss) DNA-type aptamer having a high affinity toward target antibody against MPT64 (anti-MPT64), resulting in quick inhibition of the peroxidase-like activity of MNPs via the adsorption of aptamer on the surface of MNPs. By the addition of sample solutions containing anti-MPT64, aptamer bound on the surface of MNPs would strongly interact with free anti-MPT64 and be detached from the MNPs, thereby increasing the available surface area of the MNPs and consequently yielding enhanced peroxidase activity. Using this strategy, target anti-MPT64 was successfully detected by displaying increased colorimetric intensities from the higher oxidation of employed peroxidase substrate, 2,2'-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS). Based on these results, we anticipate that aptamer adsorbed MNPs can serve as a potent probe system for the detection of clinically important target molecules.