Fluorescent biosensor for simultaneously detecting two kinds of RNA as well as preparation method and use method thereof

Abstract

The invention discloses a fluorescent biosensor for simultaneously detecting two kinds of RNA as well as a preparation method and a use method thereof. When a target detection object miRNA appears in a reaction system, the capture chains cDNA1 and cDNA2 are respectively combined with the target microRNA, and the fluorescence of the CQDs and the Cy5 is recovered. Meanwhile, double-strand specific nuclease (DSN) is used to act on the double strands formed by the pDNA and the target MicroRNA, so that the microRNA participates in the next round of reaction, and the signal is amplified. The construction method provided by the invention is simple, the detection sensitivity is high, the expression quantity of two miRNAs in a sample can be quantitatively detected at the same time, the reaction condition is mild, the sensing system is stable and effective, and compared with expensive and tedious clinical gene sequencing, the method has great practical significance and development potential and is beneficial to popularization and application.

Description

technical field [0001] The invention belongs to the technical field of biosensing, and in particular relates to a fluorescent biosensor for simultaneously detecting two RNAs and a preparation and application method thereof. Background technique [0002] At present, the commonly used detection methods for miRNA mainly include Northern blot technology, solexa sequencing technology, microarray, fluorescence in situ hybridization technology, real-time quantitative fluorescent PCR (QRT-PCR), etc., which can accurately identify the genome. Although the sensitivity is high, the operation is complicated. , time-consuming and expensive to detect. In terms of biosensors, detection methods such as electrochemical, ultraviolet, SPR, and colorimetry are often used in combination with reaction principles such as molecular beacons and enzymes, which greatly reduce detection costs and waiting time, but still have high detection limits, low sensitivity, The disadvantages are poor reproducib...

AI Technical Summary

Problems solved by technology

[0002] At present, the commonly used detection methods for miRNA mainly include Northern blot technology, solexa sequencing technology, microarray, fluorescence in situ hybridization technology, real-time quantitative fluorescent PCR (QRT-PCR), etc., which can accurately identify the genome. Although the sensitivity is high, the operation is complicated. , time-consuming and expensive

In terms of biosensors, detection methods such as electrochemical, ultraviolet, SPR, and colorimetry are often used in combination with reaction principles such as molecular beacons and enzymes, which greatly reduce detection costs and waiting time, but still have high detection limits, low sensitivity, Disadvantages such as poor reproducibility and more interference in actual samples

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