Development of carbon dot based microplate and microfluidic chip immunoassay for rapid and sensitive detection of HIV-1 p24 antigen

A highly sensitive, precisely specific, environmentally friendly, high-throughput, microwell-plate and microchip-based sandwich assay was developed to detect HIV-1 p24 antigen, a protein biomarker using fluorescent carbon dots. High quantum yield carbon dots were synthesized using citric acid and ethylenediamine as carbon and nitrogen sources by a single-step hydrothermal reaction. The desired amine groups confirmed by FTIR on the carbon dots were coupled to streptavidin by amine–amine coupling reaction using glutaraldehyde. The detection range of the carbon dot based immunoassay (CDIA) was found to be between 20 and 1000 pg/mL in a linear dose-dependent manner. CDIA tested for HIV negative plasma samples showed no false positive results in the detection of HIV-1 p24 antigen. The CDIA was extended to develop a microfluidic carbon dot immunoassay (μCDIA) which exhibited analytical sensitivity in the range of 30–1000 pg/mL. The CDIA and μCDIA can easily be adapted to a lab-on-a-chip platform for use in resource limited settings and can also be multiplexed for the detection of other pathogens like TB and Hepatitis.