Clinical diagnosis urgently requires ultrasensitive, accurate and rapid monitoring of low-abundance biomarkers. A biosensing strategy capable of detecting target genes at the femtomolar scale was designed in this work. In the biosensing strategy, the target can induce the specially designed hairpin probe H1 to self-fold and form a 3' blunt-ended structure. When there are the hybrid double-stranded P1-T1, ligase, polymerase and nickase, the target gene was recycled, and at the same time the system produces a lot of T1 and T2. T1 and T2 can simultaneously trigger HCR, causing the modified fluorophore FAM on the DNA strand to move away from the quencher group BHQ. The amplified fluorescent signal can be captured by a fluorescence instrument. It is exciting for us that three signal amplifications are involved to achieve femtomolar detection of target genes, namely target recycling, dual-triggered HCR of T1 and T2, and HCR. In addition, it still has good detection ability in actual samples simulated by serum. We expect that the sensing strategy proposed in this paper offers great potential for biomarker detection of leukemia for early clinical diagnosis.
Keywords: Fluorescence enhancement detection; Leukemia; Multistage signal amplification; Pax-5a; Target-programmed controllable signal inspiring engineering.
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