A competitive immunosensor for sensitive detection of neomycin residues in raw milk
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Abstract
Neomycin is a common broad-spectrum aminoglycoside antibiotic that inhibits the growth of both gram-negative and gram-positive bacteria. Overdose has the potential to cause ototoxicity as well as nephrotoxicity. Thus, the presence of neomycin in raw milk poses a possible health risk to consumers. Therefore, sensitive and selective methods are necessary to monitor and detect neomycin. A competitive immunosensor was designed to detect neomycin residues in raw milk. First, a microplate was coated with conjugated neomycin-bovine serum albumin and used as an analytical competitor. The competitive immunosensor was created by incubating free neomycin with anti-neomycin antibodies labelled with HRP-streptavidin. After the competitive reaction, captured HRP catalysed the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide (H2O2) to create colour responses. The proposed immunosensor had excellent analytical performance with a low detection limit of 4 ng/mL, a wide linear range from 1 to 50 ng/mL (R2 = 0.9976), high precision with a %RSD of 3.7, and high selectivity in neomycin detection compared to six competitive antibiotics. The applicability of the sensor was evaluated by analysing neomycin in raw milk samples. Detection of neomycin in real samples demonstrated remarkable accuracy, with a recovery rate of 85–111%. The results showed that the proposed immunosensor can successfully detect neomycin in raw milk samples with excellent sensitivity and ease of use. Additionally, the proposed method has great potential for farmers to detect neomycin residues in raw milk.
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