Simultaneous determination of five-type hepatitis virus antigens in 5 min using an integrated automatic electrochemical immunosensor array

An integrated automatic electrochemical immunosensor array has been designed for the simultaneous detection of 5-type hepatitis virus antigens (i.e. hepatitis A, hepatitis B, hepatitis C, hepatitis D, and hepatitis E). Initially, 5-type hepatitis virus antibodies were immobilized onto a self-made electrochemical sensor array using nanogold particles and protein A as matrices, and the immunosensor array was then used to capture their corresponding antigens from sample solution with a 1-step capture format. The detection is based on the potential change before and after the antigen-antibody reaction by using a 2-electrode system. The developed immunosensor array with allowed simultaneous determination of 5-type hepatitis virus antigens in 5 min. The detection limit of the sensor array was ≤1.0 ng/mL for most analytes. The intra- and inter-assay imprecision (CVs) of the sensor array were ≤7.3% and 8.1%, respectively. No nonspecific adsorption was evaluated, and the crossing talk between neighboring sites was ≤7.5% (CV). The developed immunoassay system offers promise for label-free, simple, rapid, cost-effective analysis of multi-analyte. Importantly, the chip-based immunosensor array could open new opportunities for high-throughput multi-analyte immunoassays in point-of-care testing of miniaturized lab-on-a-chip devices.