Immunomagnetic antibody plus aptamer pseudo-DNA nanocatenane followed by rolling circle amplication for highly-sensitive CTC detection

Abstract

Biosensing and detecting the rare circulating tumor cells (CTCs) in complex blood samples are a great challenge but necessary for cancer metastasis prevention. Here we show a novel highly-sensitive biosensing system for detecting CTCs in whole blood. The system is composed of Her2-coated immunomagnetic beads and an anti-EpCAM aptamer assembled pseudo-DNA nanocatenane (PDN) for dual targeting and separating CTCs, in conjunction with the rolling circle amplification (RCA) and molecular beacon (MB) system for CTCs signal amplification. The Her-2-coated beads separated CTCs from blood after their elution from a magnetic column. The unique PDN, which is a tailor-designed self-assembly of three circular DNAs that are inter-locked with independent and non-interfered templates for periodic RCA process, binds EpCAM-rich CTCs. In the presence of the RCA primer, phi29 DNA polymerase and MB, the system collaboratively generated the amplified fluorescent signals for highly-sensitive detection of CTCs. Through this system, we achieved the limit of detection less than 10 CTCs/mL blood, and quantified the number of CTCs in patient blood, which is proportional to the patient cancer status. Our technique is highly-sensitive, practicable and convenient enough for clinical detection of breast CTCs.