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Updated: Sep 16, 2025
Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
Published on: December 12, 2011
Ratiometric fluorescence immunoassay based on PtPdRu trimetallic nanozyme with enhanced oxidase-like activity for
Liang Luo1, Zizhe Li2, Qiang Li1
1National Key Laboratory of Veterinary Public Health and Safety, Beijing Key Laboratory of Detection Technology for Animal Derived Food Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; Technology Innovation Center for Food Safety Surveillance and Detection (Hainan), Sanya Institute of China Agricultural University, Sanya 572025, People's Republic of China.
Abstract:
Xylazine (XYL), an illicit opioid adulterant, its abuse and misuse can cause serious food safety incidents, drug-facilitated crimes, and even death. Therefore, to address the urgent need for XYL monitoring, a novel ratiometric fluorescence (RF) nanozyme-linked immunosorbent assay (NLISA) was proposed. This RF-NLISA used a robust polyvinylpyrrolidone ligand assisted trimetallic PtPdRu nanocluster (NC) nanozyme with high oxidase-like activity as the signal label. The PtPdRu NC could simultaneously catalyze the dual substrates scopoletin and Amplex Red to induce opposing fluorescence intensity changes, enabling sensitive RF signal output. Based on PtPdRu NC catalysis to trigger self-calibrating RF signals, the RF-NLISA achieved a limit detection of 0.0092 ng mL-1. This represents a nearly 10-fold sensitivity improvement over traditional horseradish peroxidase-based colorimetric ELISA, while maintaining excellent specificity and accuracy. This novel PtPdRu NC nanozyme facilitated RF-NLISA provides a sensitive and accurate sensing platform for XYL abuse monitoring in various food and human urine samples.
