Organic dyes are typically applied as photosensitizers in photoelectrochemical (PEC) cells but have not been reported in polarity-reversal-mode PEC sensors with excellent sensitivity and accuracy. Herein, an elegant and robust PEC biosensor for carcinoembryonic antigen (CEA) has been designed by photocurrent polarity switching of CdTe quantum dots (QDs), which is obtained by embedding methylene blue (MB) into amplified double-stranded DNA (dsDNA) anchored to the superparamagnetic Fe3O4@SiO2. The target-triggered Exo III-assisted cyclic amplification strategy and in situ magnetic enrichment enable the remarkable sensitivity. The extraction of target-analogue single-stranded DNA (output DNA) contributes to high selectivity resulting from the elimination of possible interferences in real samples or matrixes. Particularly, this exclusive polarity-reversal-mode PEC aptasensing can efficiently eliminate the false-positive or false-negative signals, leading to accurate measurements. Moreover, different from the probes and layer-by-layer assembled photoelectric beacons on electrodes in advance, this rational split-type approach is doomed to help the PEC biosensor with additional merits of convenient fabrication, short time consumption, wider linearity, as well as outstanding reproducibility and stability in practical applications. In light of the ability of MB acting as a kind of signal probe in typical electrochemical sensors, certainly, this ingenious design can not only be extended to a wide variety of target monitoring but also provide new ideas for the construction of high-performance electrochemical and PEC biosensors.