Method for simultaneously detecting miR-486, miR-146b and miR-15b by one-step method and application of method

Abstract

The invention discloses a method for designing miR-486, miR-146b and miR-15b probes. The method comprises the following specific step: (1) designing a sequence of stem loop-miR-486 into SH-(CH2)6-AAAAAC TCG GGG CAG CTC AGT ACA GGA TTT TT-6-FAM. According to the method, through designing three kinds of stem loop structures, different fluorescent molecules are modified by 3' ends, stem loop-miR-486is modified by FAM, stem loop-miR-146b is modified by Cy5, stem loop-miR-15b is modified by Cy3, and a gold nano surface is modified with 5' ends through gold-mercapto bonds; due to a special structure of a stem loop, the fluorescent molecules will be in contact with the gold nano surface, and fluorescence is quenched; when target miRNA is present, stem loop structures are opened due to base complementarity mating, and fluorescent signaling molecules can be far away from the gold nano surface, so that the condition that corresponding fluorescence signals are greatly increased can be detected,three kinds of miRNA of a transcription factor can be rapidly and simultaneously detected, and the sensitivity and specificity are good.

Description

technical field [0001] The present invention relates to a method for detecting miR-486, miR-146b and miR-15b and its application field, in particular to a method for simultaneously detecting miR-486, miR-146b and miR-15b in one step and its application. Background technique [0002] Type 2 diabetes (T2D) is a long-term metabolic disorder characterized by pancreatic beta. Cellular destruction and insulin resistance are major contributors to cardiovascular disease. It is well established that childhood obesity contributes to the incidence of T2D in adults. Therefore, identifying obese children at risk of developing T2D and preventing the pathological process will help reduce the incidence of T2D in adults. However, there is currently a lack of good early diagnosis methods. Our previous study found that three miRNAs, miR-486, miR-146b and miR-15b, were stably and significantly highly expressed in obese children and T2D patients. The expression levels of these three miRNAs c...

AI Technical Summary

Problems solved by technology

Using microfluidic technology and quantum dot nanoparticles with different light emission, the Fan group reported a quantum dot-based microfluidic platform to simultaneously detect two tumor markers; Wang group reported a method using magnetic nano A multi-channel microchip of particles can be used to detect multiple disease markers simultaneously; however, so far, there is no method that can rapidly and simultaneously detect miR-486, miR-146b and miR-15b

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