Novel dual-emissive fluorescent immunoassay for synchronous monitoring of okadaic acid and saxitoxin in shellfish

In this work, we present a novel dual-emissive fluoroimmunoassay for synchronous monitoring of okadaic acid (OA) and saxitoxin (STX) using multicolor fluorescent labels composed of sulfur, phosphorous co-doped graphene quantum dots (S, P-GQDs), and ovalbumin (OVA)-coated gold nanoparticles (OVA-AuNPs). The novel OVA-AuNPs were prepared by the reduction of chloroauric acid under alkaline conditions using OVA as a reducing agent. Both S, P-GQDs and OVA-AuNPs exhibit bright fluorescence, more importantly, a large emission wavelength difference (Delta lambda = 156 nm) under an excitation of 400 nm and relatively independent fluorescence behavior, which are essential to realizing the dual-signal marks in a directly mixing system. Using a competitive fluorescence-linked immunosorbent assay (cFLISA) format, the dual-emissive cFLISA was successfully utilized to measure OA and STX contents in Alectryonella plicatula (commonly named as fingerprint oyster) and the detection results were in good agreement with the commercial enzyme-linked immunosorbent assay (ELISA) kits.

Automated summary

This study presents a novel dual-emissive fluoroimmunoassay for synchronous monitoring of okadaic acid (OA) and saxitoxin (STX) using multicolor fluorescent labels. The method successfully measured OA and STX contents in Alectryonella plicatula and showed good agreement with commercial ELISA kits.