Enrichment And Detection Of MiRNA In Serum Based On Hydrogel Microfluidic System

In recent years,circulating miRNAs have entered the field of vision as a novel marker that can exist stably in the blood.miRNAs are involved in cell division and differentiation and regulation of gene expression levels,which are closely related to tumor development.However,the miRNA content in the human body is extremely small,and the humoral environment in which circulating miRNAs exist is extremely complicated.Therefore,the enrichment detection of miRNA is a key difficulty in recent research.At present,conventional enrichment detection methods such as commercially available miRNA extraction kits and commercial magnetic beads are expensive and the extraction and enrichment efficiency is not satisfactory.In this study,a set of hydrogel microfluidic system was developed to prepare agarose microdroplets containing magnetic fluid by microfluidic chip.After freezing,activating and surface-conjugated Ago2 antibody,the enriched serum can be specifically captured.Ago2-miRNA protein complex.The magnetic agarose microspheres prepared by the microfluidic method have uniform morphology,good magnetic properties,and can rapidly and efficiently enrich miRNAs in serum.The miRNA is dissociated at high temperature,the agarose microspheres are melted,and the microdroplets containing the target molecule are again generated to carry out quantitative detection of the miRNA by digital droplet PCR.The experiment is completed by the microfluidic platform,which greatly saves the amount of experimental reagents,greatly reduces the experimental cost,shortens the experimental time,improves the experimental efficiency,and isolates the outside world and reduces the pollution of the external environment.The agarose microdroplets and microspheres prepared by this platform have been experimentally proven to undergo various complex environmental changes and remain unchanged in shape,and due to the good biological activity of agarose,it can be coupled with various functional molecules.The enrichment extraction experiment can be used as a single reactor for preserving the target molecule,and various downstream processing analysis is carried out.It has broad application prospects in the fields of bioconcentration detection and single molecule single cell analysis.