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Sensitive lateral flow immunoassay of an antibiotic neomycin in foodstuffs.

ABSTRACT: Aminoglycosides belong to a class of antibiotics now widely used in agriculture and veterinary medicine and expected to contaminate food products. In this study, a sensitive lateral flow immunoassay (LFIA) of an aminoglycoside neomycin (NEO) was developed. Two methods of immunochromatographic detection based on various techniques of gold nanoparticles (AuNPs) introduction as a label were compared. It was demonstrated that the indirect labeling (a conjugation of anti-species antibodies with a marker) allowed for an increase in assay sensitivity by 80 times. The test system was characterized by an instrumental limit of detection of 0.1 ng/mL and the cutoff level of 10 ng/mL; the assay duration was 15 min. Specificity only toward NEO was demonstrated. The developed LFIA has been tested to detect NEO in different foodstuffs. It has been demonstrated that 70-119% of NEO (coefficients of variations < 10%) can be determined in milk, turkey meat, honey, and eggs using simple procedures of preliminary sample preparation. Testing the samples showed the coincidence of the results for the developed lateral flow assay and for commercial ELISA kit.

SUBMITTER: Hendrickson OD

PROVIDER: S-EPMC7813914 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description: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 (R 2 > 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.

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Sensitive lateral flow immunoassay of an antibiotic neomycin in foodstuffs.

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