A panel of miRNAs in serum extracellular vesicles serves as novel diagnostic biomarkers for MASLD.

The rising global prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) poses a growing challenge to healthcare systems, stimulating substantial research efforts to develop reliable diagnostic methodologies. Emerging evidence highlights extracellular vesicles (EVs) as promising non-invasive biomarkers due to their roles in metabolic regulation and disease progression. This study investigated the diagnostic potential of serum EV-derived microRNAs (miRNAs) for MASLD detection and staging. We developed a novel diagnostic approach combining wheat germ agglutinin (WGA)-coupled magnetic beads for EV capture with RT-qPCR analysis, creating a streamlined two-step protocol that eliminates conventional purification requirements. MiR-574-3p, miR-542-3p, and miR-200a-3p in serum EVs were significantly elevated in patients with MASLD, indicating their potential as non-invasive biomarkers. Here, our established platform offers a clinically feasible solution for EVs isolation and quantitative miRNA analysis, presenting significant advantages in diagnostic efficiency and practical implementation.