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

[Display omitted] •DNA-linked immunosorbent assay does not require the involvement of enzyme.•Aptamer-initiator strand serves as a target recognizer and signal activator.•Dual modality meets the on-spot preliminary inspection and sensitive detection.•Accuracy and sensitivity of the detection in foods are comparable to ELISA kits. 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.