Highly sensitive analysis of tetrodotoxin based on free-label fluorescence aptamer sensing system.

Tetrodotoxin (TTX) specifically can bind to its nucleic acid aptamer (TTX-aptamer) and cause the conformation of TTX-aptamer to be switched from the single-strand random coiling form to the compact neck ring structure. Based on the microenvironment difference of the fluorescence reporter, berberine in between the single-stranded coil oligonucleotides and the structure of the neck ring, a simple, rapid and sensitive label-free fluorescence aptamer sensing system for detection of TTX was developed. Various factors affecting the analysis of TTX were optimized, including the concentration of berberine, ion strength, pH, reaction time, the concentration of TTX-aptamer. Under the optimal experimental conditions, the fluorescence intensity of the sensing system and the concentration of TTX showed a good linear relationship in the range of 0.1 nM to 500 nM, with the detection limit of 0.074 nM. The standard recovery test result exhibited that the recoveries of TTX in serum samples were 96.54%-106.40%. The established method has the advantages of high specificity, good sensitivity, quickness and convenience, low cost, and can be used for the detection of TTX in serum samples.