Europium Fluorescent Nanoparticles-Based Multiplex Lateral Flow Immunoassay for Simultaneous Detection of Three Antibiotic Families Residue

Abstract

A fluorescent immunoassay based on europium nanoparticles (EuNPs-FIA) was developed for the simultaneous detection of antibiotic residues, solving the problems of single target detection and low sensitivity of traditional immunoassay methods. In the EuNPs-FIA, EuNPs were used as indictive probes by binding to anti-tetracyclines monoclonal antibodies (anti-TCs mAb), anti-sulphonamides monoclonal antibodies (anti-SAs mAb) and anti-fluoroquinolones monoclonal antibodies (anti-FQs mAb), respectively. Different artificial antigens were assigned to different regions of the nitrocellulose membrane as capture reagents. The EuNPs-FIA allowed for the simultaneous detection of three classes of antibiotics (tetracyclines, fluoroquinolones and sulphonamides) within 15 min. It enabled both the qualitative determination with the naked eye under UV light and the quantitative detection of target antibiotics by scanning the fluorescence intensity of the detection probes on the corresponding detection lines. For qualitative analysis, the cut-off values for tetracyclines (TCs), fluoroquinolones (FQs) and sulphonamides (SAs) were 3.2 ng/ml, 2.4 ng/ml and 4.0 ng/ml, respectively, which were much lower than the maximum residue limit in food. For quantitative analysis, these ranged from 0.06 to 6.85 ng/ml for TCs, 0.03–5.14 ng/ml for FQs, and 0.04–4.40 ng/ml for SAs. The linear correlation coefficients were higher than 0.97. The mean spiked recoveries ranged from 92.1 to 106.2% with relative standard deviations less than 8.75%. Among them, the three monoclonal antibodies could recognize four types of TCs, seven types of FQs and 13 types of SAs, respectively, and the detection range could cover 24 antibiotic residues with different structural formulations. The results of the detection of antibiotic residues in real samples using this method were highly correlated with those of high performance liquid chromatography (R2 > 0.98). The accuracy and precision of the EuNPs-FIA also met the requirements for quantitative analysis. These results suggested that this multiplex immunoassay method was a promising method for rapid screening of three families of antibiotic residues.

Europium Nanoparticles-Based Fluorescence Immunochromatographic Detection of Three Abused Drugs in Hair

“…According to the specific scheme of europium nano-labeled antibodies in our laboratory in the early stages and the related literature reports [ 47 ], the specific operation was as follows: 2 mg of fluorescent microspheres was added to 1 mL of MES (0.05 M, pH 5.0) solution and well mixed, centrifuged for 20 min at 12,000 rpm/min; then, the supernatant was removed and we added 1 mL MES, mixed and sonicated. We repeated the appeal operation and washed EuNPs three times.…”

Europium Nanoparticle-Based Lateral Flow Strip Biosensors for the Detection of Quinoxaline Antibiotics and Their Main Metabolites in Fish Feeds and Tissues

“…Optimization of reaction time can increase the ability of LFSB detection systems to capture and recognize target molecules, thereby improving detection sensitivity. In the study, 10 different time points (1,5,10,15,20,25,30,35,40, and 45 min) were used to obtain the optimal reaction time. The results are shown in Figure 2j.…”

Latest Trends in Lateral Flow Immunoassay (LFIA) Detection Labels and Conjugation Process

“…Europium (III) is the most widely used lanthanide for the synthesis of luminescent nanoparticles used in immunoassay and imaging applications ( Syamchand and Sony, 2015 ). Using EuNPs, Wang et al (2021) developed an LFIA for the simultaneous detection of three different types of antibiotic residues of veterinary drugs: tetracyclines, sulfonamides, and fluoroquinolones, which are common food contaminants. The analysis allowed both qualitative and quantitative detection.…”