Micro ribonucleic acid detection method based on microfluidic bead array chip and DNA polymerase-mediated primer extension technology

Abstract

The invention discloses a micro ribonucleic acid detection method based on a microfluidic bead array chip and a DNA polymerase-mediated primer extension technology. According to the detection method, a micro ribonucleic acid capture probe modified by biotin is fixed to the surface of an avidin-modified microsphere through a biotin-avidin bonding method to form a functionalized microsphere; the functionalized microsphere is fixed in a micro chamber corresponding to a detection zone of a microfluidic chip so as to form the microfluidic bead array detection chip; then a sample solution containing micro ribonucleic acid is allowed to flow into the microfluidic bead array detection chip, then to flow into a DNA polymerase solution and a biotinylation extension substrate, and finally to flow into a quantum dot labeled by the avidin; and surface fluorescence of the microsphere after the micro ribonucleic acid flows into is calculated to perform qualitative operation and quantification. The method can detect the micro ribonucleic acid with high sensitivity and high specificity and achieves quantification analysis of specificity and sensitivity of the micro ribonucleic acid in a total RNA sample.

Description

technical field [0001] The invention relates to the technical field of biological detection of a micro-total analysis system, in particular to a microRNA detection method based on a microfluidic bead array chip and a DNA polymerase-mediated primer extension technology. Background technique [0002] MicroRNA (miRNA) is a small endogenous non-coding RNA molecule, consisting of about 21-25 nucleotides, usually targeting one or more mRNAs, and inhibiting or breaking target mRNAs at the translational level. Regulates gene expression. More than 3,000 miRNAs have been discovered so far, most of which play a key regulatory role in animals and are one of the most important gene expression regulators. It is estimated that about 2 / 3 of the genes in the human body are regulated by a certain gene. Or the regulation of a set of miRNAs. Therefore, the development of new detection methods for miRNA is of great significance for medical clinical diagnosis. The developed miRNA detection met...

AI Technical Summary

Problems solved by technology

The development of this technology has better solved the problem of high-throughput and high-sensitivity detection under the condition of micro-sample. At the same time, the detection process is simple and does not require expensive detection instruments. It provides a new direction for the development of future automated high-performance biomolecular detection platform technology. , this technique has not been reported for the analysis of microRNA

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