Surface plasmon resonance imaging-based biosensor for multiplex and ultrasensitive detection of NSCLC-associated exosomal miRNAs using DNA programmed heterostructure of Au-on-Ag.

Exosomal miRNAs are potential tumor biomarkers for early diagnosis of non-small cell lung cancer (NSCLC). Herein, a surface plasmon resonance imaging (SPRi)-based biosensor was developed for simultaneous detection of multiplex NSCLC-associated exosomal miRNAs in a clinical sample using Au-on-Ag heterostructure and DNA tetrahedral framework (DTF). Exosomal miRNAs are captured by various DTF probes immobilized on the gold array chip. Subsequently, single-stranded DNA (ssDNA) functionalized silver nanocube (AgNC) hybridizes with the captured exosomal miRNAs and then the ssDNA-coated Au nanoparticles assembled on the surface of AgNC, forming Au-on-Ag heterostructures as essential labels to realize amplified SPR response. With the aid of DNA programmed Au-on-Ag heterostructure and DTF, the SPRi-based biosensor exhibits wide detection range from 2 fM to 20 nM, ultralow limit of detection of 1.68 fM, enhanced capture efficiency, and improved antifouling capability. Furthermore, the biosensor enables accurate discrimination of NSCLC patients based on detection results of exosomal miRNAs. Overall, this developed biosensor is a promising tool for multiplex exosomal miRNAs detection, providing a new possibility for early diagnosis of NSCLC.