Immunoassay based on Au-Ag bimetallic nanoclusters for colorimetric/fluorescent double biosensing of dicofol.

The high toxicity of dicofol (DICO) to nontarget organisms has resulted in the contamination of food materials and caused a threat to human health. Developing a rapid and sensitive detection method of DICO in food samples is essential and still pursued. Fluorescent nanomaterials have been widely applied in biosensors to improve the sensitivity of detection. Herein, glutathione-capped Au-Ag bimetallic nanoclusters (Au-Ag NCs) exhibited the outstanding fluorescence characteristic with the average fluorescence lifetime of 1971.08 ns and photoluminescence quantum yield of 9.84% when the molar ratio of Au to Ag was 5:1. Polyethyleneimine modified gold nanoparticles (PEI-Au NPs) with the positive charge were prepared to generate a strong colorimetric signal. A dual-model colorimetric/fluorescent immune probe based on the Au-Ag NCs and PEI-Au NPs was successfully constructed by electrostatic force, and could be applied in both ic-ELISA and LFIA methods for rapid and ultrasensitive detection of DICO. In the ic-ELISA method, the introduction of fluorescence signal significantly increased the sensitivity of detection with the limit of detection (LOD) of 0.62 ng/mL and exhibited an excellent linear relationship within the range of 1.36 ng/mL-19.92 ng/mL. In the LFIA method, the fluorescence signal of Au-Ag NCs was accumulated on the test line and control line for the fluorescence model detection with a quantitative LOD at the level of 1.59 ng/mL. Such a dual-model colorimetric/fluorescent immunoassay serves as a promising candidate to develop new approaches in field detection.