Volume 49 Number 3

Dual-targeted nucleic acid probe detection system for circulating tumor cells to enhance cancer diagnosis

Hanyun Chen^a, Yanjing Zhang^b, Haidong Dong^c, Xiujuan Zhang^d,*

 
ABSTRACT:     Circulating tumor cell (CTC)-based diagnostics represent a vital strategy for reducing cancer mortality; however, current detection methods face significant limitations including complex blood composition, the scarcity of CTCs in the bloodstream, and inconsistent expression of surface biomarkers, thereby restricting their clinical utility. Here, wepresent adual-targeted nucleic acid probe detection system (DPDS) designed for the simultaneous recognition of EpCAMandEGFRsurfacebiomarkersonCTCs. SystematicevaluationrevealedthatthisDPDSexhibitsconcentration dependent fluorescence enhancement, achieving a detection limit of 5?10 cells/ml in standard samples. When applied to blood samples from A549, MCF-7, and MDA-MB-231 tumor-bearing mice (tumor volume ?900 mm3), the DPDS demonstrated significantly improved diagnostic accuracy over single-target assays. Receiver operating characteristic (ROC)analysis yielded area under the curve (AUC) values of 0.820 (EpCAM-Cy5)and0.829(EGFR-FAM)forindividual channels, compared to a markedly superior AUC of 0.981 for the DPDS. This study develops an innovative dual-marker liquid biopsy platform that overcomes key challenges in conventional CTC detection, including low abundance and phenotypic heterogeneity. The platform demonstrates robust performance in identifying rare CTCs (5?10 cells/ml) in peripheral blood while eliminating false negatives associated with single-marker approaches, ultimately establishing a clinically viable solution for sensitive and specific cancer diagnosis.

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* Corresponding author, E-mail: jenifer81@126.com

Received 17 Jul 2025, Accepted 13 Dec 2025