Fluorescent biosensor based on nucleic acid recognition induction and application thereof

A biosensor, nucleic acid technology, applied in fluorescent biosensors and application fields, can solve the problems of low accuracy, low sensitivity, and high cost, and achieve the effects of improving accuracy, enhancing sensitivity, and enhancing strength

Inactive Publication Date: 2020-04-03
QINGDAO UNIV
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it has been proved that these methods have good sensitivity and selectivity, the inventors of the present disclosure found that these methods for detecting miRNA are slow and costly, which affects and limits their popularization and application
Due to its low cost, simple operation and good reproducibility, the fluorescence detection method can solve these problems such as slow speed and high cost. However, after further research by the inventors of the present disclosure, it is found that the fluorescence detection method has low sensitivity and high accuracy. low level problem

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fluorescent biosensor based on nucleic acid recognition induction and application thereof
  • Fluorescent biosensor based on nucleic acid recognition induction and application thereof
  • Fluorescent biosensor based on nucleic acid recognition induction and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Preparation of fluorescent probes based on gold nanoparticles (based on nucleic acid recognition-induced fluorescent biosensors):

[0041] Take 1mL gold nano solution (1mM, the appearance of gold nano is as follows figure 2 shown) in a centrifuge tube, and 20 μL of thiol-modified recognition sequence cDNA at a concentration of 100 μM was added thereto, and then the resulting mixed solution was reacted with gentle stirring at 37° C. for 12 h. In order to reduce non-specific adsorption, the cDNA-gold nanocomposites were dispersed in 200 μL 0.1 MPBS (pH 7.4) containing 3% BSA, 0.1 M NaCl and 0.02% Tween-20, kept at 37 °C for 1 h, and then washed with PBS buffer (0.1 M, pH 7.4) was washed 3 times, and redispersed in 200 μL of PBS buffer (0.1 M, pH 7.4). Next, 50 μL of two oligonucleotides (signaling probes) M1 and M2 (100 μM each) were added to the above solution, and reacted at 37° C. for 2 h. After washing three times with PBS buffer (0.1 M, pH 7.4), the resulting comp...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a fluorescent biosensor based on nucleic acid recognition induction and an application thereof. The fluorescent biosensor is composed of gold nanoparticles, a recognition sequence and n signal probes, the recognition sequence is single-stranded DNA, the surfaces of the gold nanoparticles are connected with one ends of a plurality of recognition sequences; the signal probe issingle-stranded DNA of which one end is connected with a fluorophore; and the fluorophores of the signal probes are different. The single-stranded DNA sequence of the signal probe is composed of a first DNA sequence and a second DNA sequence. The fluorophore is connected with a second DNA sequence; each recognition sequence connected with the gold nanoparticles is hybridized with the first DNA sequence of the n signal probes; the recognition sequence is complementary with the miRNA-155, and after the recognition sequence is hybridized with the miRNA-155, the 3'end of the recognition sequenceis changed into a blunt end which can be recognized and hydrolyzed by the exonuclease III, and the n signal probes are all unlinked with the recognition sequence; wherein n is a natural number greaterthan 1.

Description

technical field [0001] The disclosure belongs to the technical field of biological detection and relates to a detection method of miRNA-155, in particular to a fluorescent biosensor based on nucleic acid recognition induction and its application. Background technique [0002] The statements herein merely provide background information related to the present disclosure and may not necessarily constitute prior art. [0003] MicroRNA (miRNA), as a class of endogenous non-coding ribonucleic acid containing 18-25 nucleotides, plays an important role in physiological and pathological processes such as cell proliferation, gene expression and metabolism. Many studies have shown that the abnormal expression of miRNA is closely related to many human diseases, especially the development of cancer. There is evidence that aberrant activation of miRNA-155 in blood or cell samples can be affected by cancer. miRNA-155 is expected to be a biomarker for early diagnosis and treatment of canc...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/682C12Q1/6825
CPCC12Q1/682C12Q1/6825C12Q2521/319C12Q2563/137C12Q2565/519C12Q2563/107C12Q2525/207C12Q2565/607
Inventor 张菲菲王海燕王宗花冯明慧
Owner QINGDAO UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap