研究目的
Developing a sensitive plasmonic enzyme-linked immunosorbent assay (pELISA) method based on silver nanoprisms (AgNPRs) etching for qualitative and quantitative detection of danofloxacin (DAN), a fluoroquinolone antibiotic, in milk samples.
研究成果
The AgNPR etching pELISA demonstrated high sensitivity, good accuracy, and excellent reliability for the detection of DAN in milk, with significant improvements over traditional ELISA methods in both quantitative and qualitative detection limits.
研究不足
The study did not explore the method's applicability to other complex matrices beyond milk, nor did it extensively compare the cost and practicality of the pELISA with other detection methods in real-world settings.
1：Experimental Design and Method Selection
The study employed a plasmonic ELISA (pELISA) based on the oxidative etching of silver nanoprisms (AgNPRs) by H2O2 generated from glucose oxidase (GOx) catalysis. The method was designed for the detection of danofloxacin (DAN) using an indirect competitive assay format.
2：Sample Selection and Data Sources
Milk samples spiked with DAN were used to evaluate the method's accuracy and precision. The detection was compared with traditional ELISA using TMB as the signal transducer.
3：List of Experimental Equipment and Materials
Key materials included silver nanoprisms (AgNPRs), biotinylated monoclonal antibody (mAb), streptavidin (SA), biotinylated glucose oxidase (Biotin-GOx), and DAN-BSA conjugate. Equipment included a Varioskan LUX multimode microplate reader, LC-MS/MS system, and JEM-2100 electron microscope for TEM.
4：Experimental Procedures and Operational Workflow
The pELISA involved coating microplates with DAN-BSA, blocking, adding DAN standards and Biotin-mAb, followed by SA and Biotin-GOx. Glucose was added to generate H2O2, which etched AgNPRs, causing a color change. The process was optimized for various parameters including concentrations of Biotin-mAb, SA, Biotin-GOx, and glucose, and reaction time.
5：Data Analysis Methods
The absorbance spectra of AgNPRs were analyzed for quantitative detection, and color changes were used for qualitative assessment. The performance was evaluated based on detection limits, recovery rates, and specificity.
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