Dual-signal electrochemical sensor for detection of cancer cells by the split primer ligation-triggered catalyzed hairpin assembly

Simultaneous detection of various intracellular biomarkers is promising for early diagnosis and treatment of cancer. Herein, a split primer ligation-triggered catalyzed hairpin assembly-based on dual-signal electrochemical biosensor was constructed for the determination of two pairs of cancer mRNAs: TK1 and c-myc, survivin and GalNAc-T by using ferrocene molecular beacon and hemin molecular beacon as detection signal sources. Each pair of targets exists simultaneously, can release the split primers and ligated as the integral primers, hybridization occurred between the integral primers and part of MBs, causing a double-stranded DNA formed. The probes hybridized with the unfolded MBs and displaced integral primers. Finally, the displaced integral primers again hybridized with the MBs and initiated cycle amplification. Under the optimal conditions, the detection limit of TK1 and c-myc mRNA is as low as 0.022 nM, and that of survivin and GalNAc-T mRNA is 0.029 nM. In addition, two pairs of cancer mRNAs could act as outputs to activate an AND logic gate.