Dual-protein orthogonal extracellular vesicle sorting and NEase amplified miRNA profiling by giant magneto resistance (GMR) biochip for the diagnosis of glioma

In recent years, extracellular vesicles (EVs) have been extensively investigated as circulating biomarkers for non-invasive glioma diagnosis due to their ability to traverse the blood–brain barrier while carrying abundant biomolecules. However, the high heterogeneity of EVs in blood circulation, attributed to their diverse cellular origins, poses challenges in isolating cancer cell-secreted EVs using current sorting methods. To address this issue, aptamer-modified magnetic nanoparticles and photosensitizer-conjugated aptamer probes were employed to achieve single-step orthogonal sorting of dual-protein positive EVs within 1 h. Integrated with a nicking endonuclease-amplified giant magneto resistance (GMR) biochip, the platform enabled ultrasensitive miRNA profiling with a detection limit of 1.88 × 10 5 copy/mL. Utilizing this platform, we revealed EGFR+/CD133 + EV miRNA signatures could effectively distinguished glioma patients from healthy controls and other brain disorders with area under curve scores of 0.948 and 0.906, respectively. The system combines rapid analysis (<2.5 h), minimal sample input (50 μL plasma), and amplified detection, establishing an advanced liquid biopsy technology for neuro-oncology.