Simultaneous detection of microcysin-LR and okadaic acid using a dual fluorescence resonance energy transfer aptasensor

Algal toxins can cause neurovirulence, hepatotoxicity, and cytotoxicity in humans through the consumption of contaminated water and food. In this work, we presented a novel aptasensor for the simultaneous detection of two algal toxins, microcysin-LR (MC-LR) and okadaic acid (OA). This system employed green and red upconversion nanoparticle (UCNP) luminescence as the donors and two quenchers (BHQ(1) and BHQ(3)) as the corresponding acceptors. The two donor-acceptor couples were fabricated by hybridizing the aptamers with their corresponding complementary DNA. The results indicated that the green and red upconversion luminescence could be quenched by the quencher probes because of their highly overlapping spectrum. In the presence of MC-LR and OA, the aptamers preferred to bind to their corresponding analytes and de-hybridize with the complementary DNA. This effect became sufficiently large to prevent green and red luminescence quenching. Under the optimized experimental conditions, the relative luminescence intensity increased as the algal toxin concentrations increased, allowing for the quantification of MC-LR and OA. The relationships between the luminescence intensity and plotting logarithms of algal toxin concentrations were linear in the range from 0.1 to 50 ng mL(-1) for MC-LR and OA. As a practical application, this type of dual fluorescence resonance energy transfer (FRET) aptasensor was used to monitor the MC-LR and OA levels in naturally contaminated food samples such as fish and shrimps.