Reagent kit and method for bidirectional signal amplification detection on miRNA [micro-RNA (ribonucleic acid)]

Abstract

The invention belongs to the field of nucleic acid detection, in particularly relates to a reagent kit and a method for bidirectional signal amplification detection on miRNA [micro-RNA (ribonucleic acid)]. The reagent kit comprises first DNA (deoxyribonucleic acid) probes (1) with hairpin structures, second DNA probes (2) with hairpin structures and annular DNA probes (3). The first DNA probes with the hairpin structures can be paired with alkali bases of the miRNA, so that the hairpin structures can be opened; the second DNA probes with the hairpin structures and the miRNA can be competitively bound with the first DNA probes with the hairpin structures, so that the miRNA can be released; a DNA complex formed by the second DNA probes with the hairpin structures and the first DNA probes with the hairpin structures is provided with at least sticky end; continuous alkali groups of a section of each annular DNA probe are complementary to the corresponding sticky end of the DNA complex, and rolling cycle amplification can be carried out by the annular DNA probes and the DNA complex to obtain G quadruplex amplification products. The reagent kit and the method have the advantages that the miRNA can be measured by the aid of the reagent kit and the method, and the reagent kit is easy and convenient to operate, low in cost, high in sensitivity and good in specificity.

Description

technical field [0001] The invention relates to the field of nucleic acid detection, more specifically, to a kit and method for bidirectional signal amplification and detection of miRNA. Background technique [0002] MicroRNAs (miRNAs) are a class of short-sequence, non-coding, single-stranded small molecule RNAs with regulatory functions that exist in eukaryotic cells, with a length of about 18-24nt. miRNAs specifically inhibit or silence the expression of genes at the translational level by complementary binding to the 3' untranslated region (3'UTR) of their target mRNA molecules, so miRNAs are widely involved in various physiological processes of organisms, such as individual development, Cell proliferation, differentiation and apoptosis, regulation of metabolism and stress response, and even disease occurrence, including tumor formation. miRNAs are also involved in the regulation of important biological programs in tumor cells, indirectly function as oncogenes and tumor...

AI Technical Summary

Problems solved by technology

[0006] The traditional detection methods of miRNAs mainly include Northern blot, microarray and real-time quantitative PCR (real-time PCR). These detection methods not only requi

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