Sensitive Detection of Exosomal MiRNA for Cardiovascular Diseases with Target Initiate Proximity Ligation Assay (TIPLA)

Abstract Reliable trace exosomal miRNA detection remains challenging. Herein, we established a universal Target Initiate Proximity Ligation Assay (TIPLA), to sensitively detect miRNA by integrating target miRNA initiate cycle and proximity ligation triggered rolling circle amplification (RCA). The outstanding features of this approach is calculated as: i) three signal amplification composed by target miRNA cycle, proximity ligation initiated RCA and magnet-based enrichment ensure high-sensitive detection of exosomal miRNA with a sensitivity of 30.25 fM; ii) direct detection of miRNAs without additional reverse transcription procedure inevitable in conventional nucleic acid detection methods. This dual amplification made TIPLA a sensitive approach to detect exosomal miRNA from both culture cells and cardiovascular disease patients, revealing a potential universal molecular detection platform for the screening, diagnosis, and prognosis prediction of multiple diseases.