Poisonous mushrooms containing α-amatoxin can be lethal, making it imperative to develop a rapid and sensitive detection method for α-amatoxin. Utilizing the DNA tetrahedral structure as its foundation, the aptamer allows controlled density and orientation. Consequently, we designed aptamer tetrahedral functionalized magnetic beads that specifically target α-amanitin to release complementary DNA (C-DNA) strands. These strands were then employed as primers to initiate rolling circle amplification (RCA) with fluorescent dyes. The combination of SYBR Green I detection probes facilitated the amplification of the detection signal, enhancing the detection sensitivity of the aptasensor. The calculated detection limit was determined to be 3 ng/mL, a magnitude lower than that of other aptasensors by 2 orders of magnitude. The aptasensor integrates the advantages of high sensitivity and specificity, offering a simple and reliable rapid detection method for α-amanitin analysis.
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