Exosome surface protein and internal miRNA simultaneous detection chip and detection technology

Abstract

The invention discloses an exosome surface protein and internal miRNA simultaneous detection chip and a detection technology. The preparation method of the chip comprises the following steps: preparing a Y-shaped array chip by adopting a soft photoetching method according to a drawn mask, sequentially introducing a biotinylated gelatin solution and an avidin solution into the chip, and assemblingthe chip layer by layer; and adding avidin-modified nanoparticles to the surface of the assembled chip, and capturing an antibody CD63 to obtain the micro-fluidic chip. The invention also prepares detection antibodies modified by different quantum dots for exosome surface protein detection, and vesicles containing different molecular beacons for exosome internal miRNA detection. The method has theadvantages of low sample consumption, rapidness, simplicity and convenience in detection, good selectivity, high sensitivity, high flux and the like; in addition, the chip can realize simultaneous detection of exosome-related proteins and miRNAs, and can effectively improve the diagnosis accuracy of corresponding diseases.

Description

[0001] The invention belongs to the technical field of microfluidic chip immune detection, and specifically relates to a method for preparing a microfluidic chip for simultaneous detection of exosomal surface proteins and internal miRNAs and its application. Background technique [0002] Exosomes are vesicles about 100 nanometers in size that are actively secreted by normal and pathological cells to the outside of the cell. They are widely present in various body fluids of the human body such as peripheral blood, cerebrospinal fluid, and urine. Because the exosomes secreted by tumor cells carry the signal molecules of the original cells, the exosomes can be used as biomarkers for early detection of cancer. Compared with the free protein or miRNA in the blood, the protein or miRNA carried by the exosomes produced by cancer cells is more stable and has more tissue specificity. Therefore, the exosomes have greater advantages as a diagnostic biomarker. However, the content of specifi...

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to solve the problem that the existing exosome detection methods require complex sample pretreatment, and cannot detect various proteins on the surface of exosomes or internal miRNA with high throughput and high sensitivity. Microfluidic chip preparation method for simultaneous detection of exosome surface protein and internal miRNA, so as to achieve efficient capture of exosomes in serum, rapid, accurate and highly sensitive quantitative detection of different markers

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