Abstract
Extracellular vesicles (EVs) have emerged as valuable biomarkers in liquid biopsies owing to their stability, accessibility, and ability to encapsulate nucleic acids. The majority of existing methodologies for detecting EV nucleic acid biomarkers require the lysis of EVs to extract DNA or RNA. This process is labor-intensive and may lead to the loss and degradation of nucleic acids. However, the emerging field of in situ EV assays offers innovative tools for liquid biopsy, facilitating direct profiling of nucleic acids within intact EVs and reducing sample handling procedures. This review focuses on the promising and innovative field of in situ EV nucleic acid analysis. It examines the translational potential of in situ EV nucleic acid analysis in liquid biopsies from detection strategies, diagnostic applications, and diagnostic aids for single EV analysis and machine learning techniques. We highlight the innovative approach of in situ EV nucleic acid assays and provide novel insights into advancing liquid biopsy technology. This approach shows a promising avenue for improving EV-based cancer diagnosis and guiding personalized treatment with minimal invasiveness.
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