The invention belongs to the technical field of biosensors, and relates to a preparation method and application of an
electrochemical biosensor based on
DNA walker. The method comprises the followingsteps: firstly, mixing
a DNA walkerAFB1
aptamer with hairpin
DNA1 and hairpin
DNA2, dropwise adding a mixture to the surface of a gold
electrode, performing reacting, dropwise adding MCH, and performing incubating at normal temperature to obtain an
electrochemical biosensor; then, modifying the surface of the
biosensor with AFB1, anchoring a
mixed solution of a probe 1, short-chain DNA1 and short-chain DNA 2, soaking the
biosensor into a
methylene blue solution, and constructing a
standard curve by measuring the current value of a sensor and the concentration of the AFB1; and finally, measuring the current value of an unknown sample, and substituting the current value into the constructed
standard curve so as to achieve the detection of the AFB1 in the unknown sample. According to the invention, dual
signal amplification can be carried out by using an extremely small amount of target AFB1 to obtain a larger
methylene blue electrochemical
signal, and the method has good detection selectivity, high speed and high sensitivity.