Smartphone-based competitive immunoassay for quantitative on-site detection of meat adulteration

Rapid, sensitive, and portable analytical methods for on-site inspection of food fraud are now an urgent requirement to ensure food quality and satisfy the ethnic considerations of consumers. Hence, for the first time, a colorimetric smartphone-based immunoassay was developed for the on-site detection of pork adulteration in meat. In detail, the immunoassay was based on a competitive strategy in which immobilized standard porcine IgG competed with the target porcine IgG extracted in a single step from meat samples. The parameters involved in each step of the immunoassay conception and the digital colorimetric detection were carefully investigated and optimized. Using polystyrene microplates as ready-to-use stable and portable immunoplatforms, TMB as chromogenic substrate, smartphone as signal readout, and Image J software for image processing; the developed immunoassay was able to detect as low as 0.01% of pork in meat mixtures in a total assay time of 30 min. The selectivity of the immunoassay was evaluated for different meat species, and it was shown to selectively respond only to pork. Furthermore, excellent stability of the prepared immunological platform was demonstrated under extreme temperature conditions (50 °C), which confirms its high portability potential for in situ quantification of pork, while being relatively cost effective and non-laborious. The developed method also provides great precision (RSD < 6%) and accuracy (relative error< 6%). Given the universal use of smartphones as portable and affordable devices, such format of immunoassay could be a promising approach for rapid and sensitive real-time monitoring of food fraud.