Biomics

An actual task is to develop a quick and affordable way to quantify the residual amounts of pollutants in food, e.g. antibiotics in honey. A DNA-aptasensor was created to determine the concentration of oxytetracycline in aqueous solutions in range from 1 to 100 ppm using a screen printed electrode and affordable electronic components. The aptamers were obtained by systematic evolution of ligands by exponential enrichment (SELEX) from a library of sequences containing 50 random nucleotides flanked by 20-nucleotide sequences for primer annealing. To perform SELEX, the oxytetracycline was immobilized on the microparticles of 40% polyacrylamide gel. There were performed 5 rounds of SELEX, including DNA annealing on immobilized oxytetracycline, washing microparticles of unbound DNA by centrifugation, elution of bound DNA by heating and subsequent PCR. Aptamers containing aminolink at the 5’end were immobilized on the screen surface of printed electrode. The direct nonlinear relationship between the aptasensor response and the oxytetracycline concentration in the range of 1-100 ppm with a sensitivity of 2.5 to 0.02 µS/ppm was shown by conductometry. The method of cyclic voltammetry shows the linear relationship between anode peak value and decimal logarithm of the oxytetracycline concentration in the range of 1-100 ppm.

determination of antibiotic concentration; oxytetracycline; biosensors; aptamers; SELEX

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