DNA-Driven Photothermal Amplification Transducer for Highly Sensitive Visual Determination of Extracellular Vesicles

Extracellular vesicles (EVs) are crucial focus of current biomedical research and future medical diagnosis. However, the requirement for specialized sophisticated instruments for quantitative readouts has limited the sensitive measurement of EVs to specialized laboratory settings, which in turn has limited bench-to-bedside translation of EV-based liquid biopsies. In this work, a straightforward temperature-output platform based on a DNA-driven photothermal amplification transducer was developed for the highly sensitive visual detection of EVs using a simple household thermometer. The EVs were specifically recognized by the antibody-aptamer sandwich immune-configuration that was constructed on portable microplates. Via a one-pot reaction, cutting-mediated exponential rolling circle amplification was initiated in situ on the EV surface, generating substantial G-quadruplex-DNA-hemin conjugates. Significant amplification in temperature was achieved from the effective photothermal conversion and regulation guided by the G-quadruplex-DNA-hemin conjugates in the 3,3',5,5'-tetramethylbenzidine-H₂O₂ system. Through obvious temperature outputs, the DNA-driven photothermal transducer enabled highly sensitive EV detection at close to the single-particle level and supported the highly specific identification of tumor-derived EVs directly in serum samples, without the requirement of any sophisticated instrument or labeling process. Benefiting from highly sensitive visual quantification, an easy-to-use readout, and portable detection, this photothermometric strategy is expected to be deliverable across professional on-site screening to home self-testing as EV-based liquid biopsies.