Novel functional DNA-linked immunosorbent assay for aflatoxin B1 with dual-modality based on hybrid chain reaction

Abstract

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins, which is frequently detected in agricultural products. Herein, a novel functional DNA -linked immunosorbent assay (DLISA) with dual-modality based on hybrid chain reaction (HCR) has been successfully developed for ultrasensitive detection of AFB1. The strategy relies on AFB1 immune-bridged occurrence of HCR and the salt-induced aggregation of gold nanoparticles (AuNPs). An aptamer-initiator stand (Apt-Ini stand) is designed for the AFB1 recognition and the activation of HCR, which can recognize the matched hairpins and cause the crossing-opening of H1 and H2, producing a long double-stranded DNA polymer. The addition of SYBR Green I achieves the fluorescent signal output. Remaining less DNA hairpins were added and stuck on the surface of AuNPs, which were insufficient to protect the AuNPs, resulting in the salt-induced aggregation with the color change from red to blue. The dual-modality provides limits of detections of 1.333 × 10−14 g/mL and 2.471 × 10−15 g/mL, respectively. This DLISA with dual-modality provides not only a colorimetry that can meet the needs of on-the-spot preliminary inspection, but also a fluorescence assay that can acquire the precise results.