Enzyme-powered cascade three-dimensional DNA machine for the ultrasensitive determination of kanamycin

DNA walking machines have been widely used in rapid and sensitive detection. In this work, we develop a single enzyme-powered DNA cascade machine for the ultrasensitive determination of kanamycin. To construct the cascade manner, two types of single-legged three-dimensional DNA walking machine are employed to implement integrated target recognition, signal transduction and signal amplification. Upon adding kanamycin to trigger the upstream machine, the sequential enzymatic cleavage drives the autonomous movement of the walking strand and produces plenty of dye-labeled fragments with fluorescence recovery. Meanwhile, these fragments also serve as walking strands to activate the downstream machine for cascade signal amplification. Taking advantage of this cascade DNA machine, ultrasensitive determination can be accomplished in 60 min. Under the optimum conditions, this method was highly selective toward kanamycin with a detection limit of 28 fM. This cascade signal amplification shows great potential for the rapid screening of antibiotics in food.