Rapid Colorimetric Assay of Tetracycline Antibiotics based on Label-free Aptamer-gold Nanoparticle (AuNPs) Probes
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Abstract:
A label-less, simple, and rapid colorimetric assay for residual tetracyclines (TCs) was developed by combining aptamer probes with gold nanoparticles (AuNPs) as the signal transduction element. This method was used to rapidly detect residual TCs in food under optimal conditions. Additionally, this novel method was then compared to the national standard method for TC residue detection (GB 31658.6-2021). The samples were extracted by methanol and acetonitrile at a 6:4 ratio (V/V) before being assayed using the proposed colorimetric assay. Given an aptamer concentration of 150 nmol/L, NaCl concentration of 40 mmol/L, and reaction time of 5 min, the linear detection range of our colorimetric method for four types of TC was 0.25~5.0 ng/mL (R>0.981). The visual detection limit was 0.5~1.0 ng/mL, and the detection time was approximately 5 min. This method is specific for tetracycline, oxytetracycline, chlortetracycline, and doxycycline, as it has no cross-reactivity for gentamicin and chloramphenicol. Spike recoveries of 63.52%~102.48% and relative standard deviations of 2.49%~6.81% were obtained when detecting TCs in cow’s milk. Therefore, the proposed colorimetric assay is simple, fast, cost efficient, and highly sensitive, which makes it suitable for the rapid batch-screening of TCs in food.
