Abstract
A duplex surface enhanced Raman scattering (SERS)-based lateral flow immunosensor was established for the simultaneous detection of two common antibiotic residues including tetracycline and penicillin in milk. The newly synthesized Au@Ag nanoparticles were labeled with different Raman molecules including 5,5-dithiobis-2-nitrobenzoic acid (DTNB) or 4-mercaptobenzoic acid (MBA), followed by the conjugation of anti-tetracycline monoclonal antibody or anti-penicillin receptor, forming two kinds of SERS nanoprobes. The two nanoprobes can recognize tetracycline-BSA and ampicillin-BSA, respectively, which facilitates the simultaneous detection of the two types of antibiotics on a single test line. After optimization, detection limits of tetracycline and penicillin as low as 0.015 ng/mL and 0.010 ng/mL, respectively, were achieved. These values were far below those of most of other documented bio-analytical approaches. Moreover, the spiking test demonstrates an excellent assay accuracy with recoveries of 88.8% to 111.3%, and satisfactory assay precision with relative standard deviation below 16%. Consequently, the results demonstrate that the SERS-based lateral flow immunosensor developed in this study has the advantages of excellent assay sensitivity and remarkable multiplexing capability, thus it will have great application potential in food safety monitoring.
Highlights
Antibiotics such as tetracycline and penicillin are frequently used to prevent and treat cow mastitis [1]
If the sample solution did not contain tetracycline, penicillin, and related analogs, the surface enhanced Raman scattering (SERS) nanoprobes could bind with the capture antigens on the test line to form a visible test line
If the sample solution contained tetracycline or penicillin molecules, the antibiotics in the solution can first bind with the SERS nanoprobe in the micro-plate well, and less free SERS nanoprobes would react with the capture antigen on the nitrocellulose membrane, and less SERS nanoprobes would be captured by the coating antigen
Summary
Antibiotics such as tetracycline and penicillin are frequently used to prevent and treat cow mastitis [1]. Over-dose use or inadequate withdrawal time can lead to antibiotic residues in milk and cause potential health hazards to consumers [2]. Previous studies indicated that consumption of antibiotic retained milk would pose the risks of allergies, hepatotoxicity, nephrotoxicity, and other direct toxicities to human beings [3]. To prevent the health risk, the authorities have set the maximum residue limits (MRLs) of antibiotic residue in milk [4]. Antibiotic residue may lead to bacterial resistance and the predicted-no-effect-concentrations (PNECs) for resistance selection. For ensuring consumer health, it is very necessary to monitor the low-level antibiotic residue in milk
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