Multiplex biosensing platform for simultaneous and precise evaluation of dual tuberculosis biomarkers.

A multiplex electrochemiluminescence (ECL) biosensing platform utilizing a potential-resolved pattern was constructed for precise evaluation of dual tuberculosis (TB) biomarkers, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). In the study, the synthesized carbon quantum dot (CQD) and gold nanocluster (AuNC) were successively combined on gold nanoparticle (AuNP) and magnetic bead (MB) to construct signal-amplifying and potential-resolved ECL complex probes. Primary antibody (Ab1) to IFN-γ and TNF-α was modified onto double distinct regions of patterned indium tin oxide electrodes for capturing TB biomarkers. Then, the captured IFN-γ and TNF-α was recognized by corresponding secondary antibody (Ab2)-functionalized ECL complex probes. The binding events generated two distinct and well-resolved ECL peaks in a single potential scan. The intensity of ECL peaks correlated linearly with IFN-γ and TNF-α concentrations over the range of 0.01-1500 pg mL. The multiplex biosensing platform enabled simultaneous and precise evaluation of dual TB biomarkers in serum, potentially establishing a faster and more reliable TB diagnostic technology.