Probe, kit and method for cascade amplification testing of miRNAs based on coding suspension microchip

Abstract

The invention discloses a probe, a kit and a method for cascade amplification testing of miRNAs based on a coding suspension microchip. The method comprises the following steps: with a target miRNAs as a template, performing a connection reaction on the template, a padlock probe and an RNA ligase in a connecting buffer solution to realize cyclization of the padlock probe; mixing the padlock probe,cyclized through connection, with a rolling circle amplification primer, a polymerase, an endonuclease and other components required by isothermal cascade amplification, and performing an isothermalcascade amplification reaction; performing a base stacking hybridization reaction on a product obtained through an isothermal cascade amplification reaction, a coding suspension microchip coupled witha capture probe, and a generalized label in a hybridization buffer solution; and after the base stacking hybridization reaction is completed, observing the coding suspension microchip by an optical detection device to realize detection of the target miRNAs. The probe, the kit and the method can realize low-cost, high-sensitivity and high-throughput detection of detected samples.

Description

technical field [0001] The invention relates to a method for detecting microRNAs (miRNAs), in particular to a method, probe, kit, system, etc. for detecting microRNAs based on cascade amplification of coded suspension microchips. Background technique [0002] MicroRNAs (miRNAs) are small endogenous non-coding RNAs that can bind to the 3' untranscribed region (3'UTR) of messenger RNA (messengerRNA, mRNA) and inversely regulate gene expression levels at the post-transcriptional stage . Current research shows that miRNAs form a complex and highly ordered regulatory network in organisms, and almost participate in every process of normal life activities. Therefore, the analysis of miRNAs can not only be used to understand its regulatory network, but also has important value and significance in clinical application. However, miRNAs have short fragments (only 19-25 bases), low abundance in cells, and very highly homologous similar sequences. These characteristics have brought ser...

AI Technical Summary

Problems solved by technology

Conventional nucleic acid amplification techniques often require complex and sophisticated sequence design to meet the requirements of multiple analysis of miRNAs, which also limits their wide application in the flux analysis of miRNAs.

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