A fluorescent assay for sensitive detection of kanamycin by split aptamers and DNA-based copper/silver nanoclusters

Kanamycin was a group of essential antibiotics generally served in treating infections of animals which leached into the environment residual in food, causing health concerns. Thus, selective and sensitive monitoring of kanamycin was significant for food safety. In this work, split aptamers were used as templates to prepare fluorescent Cu/Ag NCs for detection of kanamycin. According to the impressive affinity of the aptamer to kanamycin, two different detection modes were designed using kanamycin aptamer as a recognition molecule, in which one was to combine split aptamer Apt-1 with Apt-2 to form an entangled DNA as a Cu/Ag NCs template, the other was to associate the normal aptamer after encirclement to form Cu/Ag NCs templates. After the addition of kanamycin, the fluorescence signals of the Cu/Ag NCs synthesized in the two modes were both enhanced, but the approach with split aptamer exhibited a superior observable sensitivity than that of the normal type. The detection range showed a well linear relationship between 80 nM and 10 μM when the emission wavelength was 560 nm, and the detection limit was 13.3 nM. In addition, when streptomycin, oxytetracycline, chloramphenicol and chlortetracycline were involved in the selective interference experiment under the same conditions, the fluorescence intensity of the system performed no significant changes. The results demonstrated that this method possessed favorable specificity and selectivity for the assay of kanamycin, proficiently achieving efficient, rapid and sensitive evaluation of kanamycin in the milk samples.