Microfluidic-Integrated Multicolor Immunosensor for Visual Detection of HIV-1 p24 Antigen with the Naked Eye

Abstract

Here, a fully integrated multicolor immunosensor was developed for sensitive and reliable semiquantitative analysis of HIV-1 p24, which integrates the multistep reactions of horseradish peroxidase (HRP)-linked immunoassay and gold nanorod (AuNR)-based multicolor assay into a single microfluidic chip. The HRP-linked immunoassay functions by moving magnetic beads bound to a capture antibody through different aqueous phases containing immunoassay reagents. HRP-catalyzed oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) is used to mediate AuNRs etching for producing various color changes. Multiple etching processes can be activated by simple mixing of the reagents from the reagent storage reservoir. The fully integrated strategy with sample-in answer-out capability is initiated by simple chip manipulation and finally concluded by converting recognition of antigen-antibody into a vivid color variation for direct visualization and semiquantitative analysis. By bare eye observation, our integrated multicolor immunosensor allows sensitive and reliable semiquantitative analysis of HIV-1 p24 within 1 h. The microfluidic chip device demonstrated here simplifies the operation significantly and thus allows broader application of a multicolor immunosensor for point of care (POC) testing in low-resource settings.