A competitive, bead-based assay combined with microfluidics for multiplexed toxin detection

Abstract

The requirement for rapid, in-field detection of cyanotoxins in water resources necessitates the developing of an easy-to-use and miniaturized system for their detection. We present a novel bead-based, competitive fluorescence assay for multiplexed detection of two types of toxins: microcystin-LR (MC-LR) and okadaic acid (OA). To automate the detection process, a reusable microfluidic device, termed toxin-chip, was designed and validated. The toxin-chip consists of a micromixer where the target toxins were efficiently mixed with a reagent solution, and a detection chamber for magnetic retainment of beads for downstream analysis. Quantum dots (QDs) were used as the reporter molecules to enhance the sensitivity of the assay and the emitted fluorescence signal from QDs was reversely proportional to the amount of toxins in the solution. An image analysis program was also developed to further automate the detection and analysis steps. Two toxins were simultaneously analyzed on a single microfluidic chip, and the device exhibited a low detection limit of 10⁻⁴ μg ml⁻¹ for MC-LR and 4 × 10⁻⁵ μg ml⁻¹ for OA detection. The bead-based, competitive assay also showed remarkable chemical specificity against potential interfering toxins. We also validated the device performance using natural lake water samples from Sunfish Lake of Waterloo. The toxin-chip holds promise as a versatile and simple quantification tool for cyanotoxin detection, with the potential of detecting more toxins.