Testing method and kit for secondary circulation amplification of microRNA (Ribose Nucleic Acid)

Abstract

The invention provides a testing method and a kit for secondary circulation amplification of microRNA (Ribose Nucleic Acid). According to the detection method provided by the invention, a secondary circulation amplification reaction can be finished by only a single operation, the sensitivity is high, and nine RNA chains in a 15-microliter reaction system can be detected; the design is simple, the compounding and modifying of nanometer materials are not related; the noise is low, so that the sensitivity, specificity and generality are increased; compared with a conventional Northern blot assay method and a conventional microarray assay method, the required sample amount is little, and the specificity is excellent; compared with a real-time quantitative PRC (Polymerase Chain Reaction), the method is a thermostatic reaction, so that the test can be realized on a normal fluorescence instrument, the use range of the method is enlarged, and the sequence design is simple and convenient; and the miRNA in a level of a single breast cancer cell can be detected. The technique disclosed by the invention can be used for designing diagnostic kits and devices for tumor.

Description

technical field [0001] The invention designs a single-molecule-level secondary cycle amplification technology to realize rapid and one-step detection of microRNAs (miRNAs) and a kit thereof. Background technique [0002] MiRNAs are a large family of highly conserved endogenous non-coding RNAs with a length of about 18-22 nucleotides, which widely exist in eukaryotes. Because miRNAs can bind to target messenger RNA through base complementarity, resulting in its degradation and inhibition of protein synthesis, thereby regulating the level of post-transcriptional gene expression. miRNAs have a very wide range of gene regulation and expression functions, and play an important role in the growth, early development, cell differentiation, and apoptosis of organisms. The expression in normal tissues and tumor tissues is significantly different, and they are involved in the occurrence of tumors ,develop. [0003] Northern blot and gene chip are the most widely used miRNAs detection...

AI Technical Summary

Problems solved by technology

However, his realization requires precise temperature cycle control, which seriously limits his further promotion, and because the sequence of miRNAs is very short, the design of PCR becomes very complicated.

Recently, some research groups have proposed to use nicking endonuclease or exonuclease to achieve constant temperature amplification reaction of nucleic acid, but they either have certain requirements for the sequence of the detected nucleic acid or the sensitivity is not high

Images