Simultaneous detection of membrane protein and mRNA at single extracellular vesicle level by droplet microfluidics for cancer diagnosis

作者：Huixian Lin, Bo Li, Jingyun Guo, Xueying Mai, Haiyang Yu, Weilun Pan, Bodeng Wu, Wei Liu, Mingzhen Zhong, Tong Liao, Ye Zhang, Bo Situ, Xiaohui Yan, Yifan Liu, Chunchen Liu, Lei Zheng · 发表于：Journal of Advanced Research · 年份：2024 · DOI：10.1016/j.jare.2024.08.026 · 被引用次数：32 · 研究领域：Extracellular vesicles in disease、Innovative Microfluidic and Catalytic Techniques Innovation、Advanced Biosensing Techniques and Applications

A new droplet microfluidic assay was proposed for the simultaneous detection of membrane proteins and mRNA at the single EV level, achieving high sensitivity in EVs subpopulation analysis. By quantitative identification of multiple EVs subpopulations, this method enabling the extraction of more tumor-related information and improving cancer diagnostic performance. It shows great potential in cancer diagnosis and could be extended into the investigation of other diseases. • A versatile droplet digital analysis assay was developed for simultaneous detection of membrane protein and mRNA at single EV level. • This assay has integrated the single-molecule sensitivity of droplet microfluidics with the high specificity of antigen–antibody reactions facilitated by antibody-DNA conjugates. • The simultaneous detection of protein and mRNA at the single EV level not only addresses the challenge of EV heterogeneity but also simplifies the detection procedure enhancing the fidelity of tumor message detection. • This study proposes a promising strategy for accurate identification and analysis of specific EV subgroups through the co-detection of proteins and mRNA at the single EV level enabling the extraction of more tumor-related information and improving cancer diagnostic performance. It shows great potential in cancer diagnosis and could be extended into the investigation of other diseases. Simultaneous detection of proteins and mRNA within a single extracellular vesicle (EV) enables com...