Single-EV MagaDecoder: Combining extracellular vesicles magnetic enrichment and multidimensional biomarkers digital decoding by droplet microfluidics for gastric cancer diagnosis and monitoring.

The information regarding tumor cells, the immune environment and microorganisms can provide insights into the formation and progression of gastric cancer (GC) from various perspectives. However, there remains a need for a diagnostic strategy capable of integrating and analyzing these multiple dimensions to accurately assess the progression of GC. Extracellular vesicles (EVs), which carry multidimensional information, represent a promising non-invasive biomarker for the diagnosis and monitoring of GC. Despite their potential, the development of an efficient method for the enrichment and precise analysis of individual EVs remains a significant challenge. Herein, we present a versatile droplet digital assay, termed the Single-EV MagaDecoder, designed to enable both the enrichment of EVs and the decoding of individual EVs. This assay consisted of two main components: efficient reversible magnetic enrichment of EVs and single EV multidimensional biomarker decoding analysis. To evaluate the clinical utility of the single-EV MagaDecoder for GC diagnosis, screening (n = 64) and validation (n = 79) GC cohorts were utilized. This assay demonstrated exceptional performance in predicting gastric disease risk, achieving 100% accuracy, and distinguishing between benign and malignant conditions with an area under the curve (AUC) of 0.8875. These results were obtained through the detection of multidimensional signatures of GC-EVs in a validation cohort (n = 79). In summary, this study proposes a promising strategy for the precise analysis of multidimensional EV subpopulations at single EV level, thereby advancing the field of precision medicine for GC.