iMEMQ: Sensitive and Versatile Detection for Extracellular Vesicles via Immunomagnetic EVs’ mRNA qPCR

Extracellular vesicles (EVs) are critical carriers of various biomolecules and present considerable potential for diagnostic and therapeutic applications. However, significant challenges remain due to the lack of straightforward and efficient methods with minimal technical barriers for the enrichment and analysis of EVs. This study presents a pioneering systematic investigation into the distribution characteristics of various nucleic acid molecules within EVs, demonstrating that internal mRNA, but not external DNA, can serve as ideal markers. By integrating upstream immunomagnetic bead capture, immunomagnetic EVs' mRNA quantitative polymerase chain reaction (iMEMQ) was established, which provides a comprehensive process for EV enrichment and analysis, with up to 260.0-fold greater detection sensitivity than that of ELISA, 53.9-fold greater sensitivity than that of immuno-PCR for CD9, and 775.6-fold greater sensitivity than that of ELISA for EphA2 in EVs. The iMEMQ ensures high-efficiency enrichment and high-sensitivity detection for different EV subgroups, providing a flexible and accessible strategy for applying EVs in fundamental research and potential disease diagnosis.