Sensitive Multicolor Visual Detection of Exosomes via Dual Signal Amplification Strategy of Enzyme-Catalyzed Metallization of Au Nanorods and Hybridization Chain Reaction.

Exosomes as nanosized vesicles have been recognized as potential noninvasive biomarkers for early cancer diagnosis. Herein, we presented a sensitive multicolor visual method for exosome detection based on enzyme-induced silver deposition on gold nanorods (Au NRs). To achieve highly sensitive determination of exosomes, hybridization chain reaction (HCR) was employed to introduce more alkaline phosphatase (ALP) for signal amplification. First, exosomes were captured by magnetic bead-labeled CD63 aptamer, and, then, cholesterol-modified DNA probes were spontaneously inserted into the exosomal lipid membrane. The ends of the DNA probes act as the initiator to trigger the HCR for signal amplification. Finally, with the help of HCR, increased sites led to enhanced ALP loading and thus boosted the ascorbic acid generation. Silver ions were reduced by ascorbic acid, and silver shells were formed on Au NRs, giving rise to the blue shift of the longitudinal localized surface plasmon resonance peak. Correspondingly, the concentration of exosomes can be obviously distinguished with naked eyes via the vivid color variation. Due to the dual signal amplification of HCR and metallization of Au NRs, highly sensitive detection for exosomes were realized with detection limits as low as 1.6 × 102 particles/μL by UV-vis spectroscopy and 9 × 103 particles/μL by naked eyes. Compared to the reported colorimetric methods for exosome quantification, visualization based on plentiful color tonalities is the most captivating merit of our approach, and HCR-induced signal amplification highlights the virtue of the strategy. The applicability of the method was validated by the analysis of clinical samples.