ATP bioluminescence-based method for screening of multiclass antibiotic residues in animal derived foods

Abstract

In present study, a new ATP bioluminescence assay with Geobacillus stearothermophilus ATCC12980 as an indicator bacterium was developed to screen multiclass antibiotic residues in animal derived foods. The development experiments includes the optimization on the usage of D-luciferin, bacterial ATP extraction condition, incubation condition of detection system and sample pre-treatment. Additionally, the validation experiments of sensitivity, specificity, accuracy, precision and stability were performed. This method can detect β-lactams (6), aminoglycosides (4), tetracyclines (4), sulfonamides (4), macrolides (4), lincosamides (1) and quinolones (3) in muscle and kidney of swine at or close to maximum residue limits (MRLs) established by European Union (EU). The false-positive rate was 0%, and the false negative rates were < 5%. The recovery rate of antibacterial drugs were between 60% and 120%, and the daily coefficients of variation were less than 15%, which meets the criteria in Commission Decision 2002/657/EC. The shelf life of the detection system was more than 5 months. Accordingly, the proposed ATP bioluminescence assay shows great potential for rapid, sensitive and high throughput detection of multiple antibiotic residues in animal foods, which can provide a preliminary screening informations of antibiotic residues for the following chemical confirmation experiments and expedite disposal decisions of unqualified food for enhanced food safety inspection.