Smartphone assisted immunodetection of HIV p24 antigen using reusable, centrifugal microchannel array chip

Abstract

An integrated immunodetection platform employing a simple, reusable, centrifugal microchannel array chip and a smartphone as detection unit was developed. The applicability of the platform to the detection of HIV p24 antigen was demonstrated. The microchip was made of polycarbonate and contained 4 × 8 zigzag microchannels. After the monoclonal antibody of HIV p24 was adsorbed onto the channel surfaces, HIV p24 was introduced into the microchannel to react with the antibody. A biotin linked polyclonal antibody was then brought in to react with HIV p24, and SP80 (containing streptavidin and horseradish peroxidase) was introduced to react with the biotin. Finally, a solution containing 3,3′,5,5′-tetramethylbenzidine and other reagents was passed through the above channels, horseradish peroxidase catalyzed the oxidation of tetramethylbenzidine (to 3,3′,5,5′- tetramethylbenzidine diamine) forming a dark color. The color intensity, indicating HIV p24 antigen quantity, was then photographed via a smartphone, and the color of each microchannel was processed via a computer to determine the HIV p24 antigen concentration. Under the optimized conditions, limits of detection (LODs) of 0.17 ng/ml and 0.11 ng/ml were obtained for p24 antigen in a buffer solution and human serum, respectively. Channel washing/rinsing was implemented via a centrifugal force. An economic portable centrifugal device that could accommodate up to 4 microchips was assembled, and multi-step solution loading and rinsing involved in this sandwich immunoassay were performed conveniently. The microchip could be reused after a simple regeneration process. The low-cost polycarbonate microchip and centrifugal device together with the simple but efficient operation make the method a promising tool for HIV screening in resource limited areas.