Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Detection of Adenosine by Label-free Fluorescent Aptamer Sensor Based on Dual Amplification Strategy

An aptamer sensor and label-free technology, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of reducing the binding constant of the aptamer itself, affecting the binding of the aptamer to the target, etc.

Active Publication Date: 2017-07-25
SHANDONG UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the invention divides the aptamer chain that recognizes adenosine into two parts, this reduces the binding constant of the aptamer itself and affects the binding of the aptamer to the target

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
  • Detection of Adenosine by Label-free Fluorescent Aptamer Sensor Based on Dual Amplification Strategy
  • Detection of Adenosine by Label-free Fluorescent Aptamer Sensor Based on Dual Amplification Strategy
  • Detection of Adenosine by Label-free Fluorescent Aptamer Sensor Based on Dual Amplification Strategy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 A label-free fluorescent aptasensor constructed based on a dual amplification strategy was used to detect adenosine.

[0070] 1. Materials and Reagents

[0071] All nucleic acid probes used in this example were synthesized and purified by Sangon Bioengineering (Shanghai) Co., Ltd., and the base sequences are shown in Table 1. The DNA stock solution was TE Buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA-2Na), and it was stored at -20°C. Adenosine was purchased from Sigma-Aldrich (Shanghai, China). Cytidine, uridine, guanosine, Exo III and DMSO were all purchased from Shanghai Sangong. Streptavidinized magnetic spheres (350 nm in diameter, 0.05% Tween-20, and 10 μM EDTA at a concentration of 3.324×10 11 beads mL -1 ,1.343g mL -1), Tris (>99.8%) was purchased from Amresco Inc. NMM (N-methylmesoporphyrinIX) was purchased from J&K Scientific Ltd. (Beijing, China). The NMM stock solution was dissolved in DMSO and stored at -20°C in the dark. All reagents (analytic...

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a label-free fluorescence aptamer sensor detection adenosine based on double-amplification strategy construction. A method comprises the following steps: (1), preparation of a streptavidin magnetic sphere probe; (2), DNA circulation and HCR reaction assisted by EXo III; (3), fluorescence measurement: after the HCR reaction is completed, adding NMM and KCl to reaction products, and carrying out fluorescence measurement after incubation to obtain a fluorescence value; (4), calculation: calculating the concentration of adenosine, in an object to be detected, containing the adenosine by utilizing the following linear equation according to the measured fluorescence value. The combination of two signal amplification strategies provides higher sensitivity for the detection of low adenosine level in a body fluid; the method can also prominently distinguish the content of adenosine in the urine of a cancer patient with the content of adenosine in the urine of a normal person, and the method can provide a novel reliable strategy for quantifying the adenosine for medical research and early clinical diagnosis.

Description

technical field [0001] The invention relates to a label-free fluorescent aptamer sensor for detecting adenosine, which is constructed based on the double amplification strategy of exonuclease III-assisted DNA circulation and hybridization chain reaction. Background technique [0002] Adenosine is an endogenous nucleoside and a degradation product of ATP, which plays very important roles in the central nervous system, peripheral nervous system, and immune system [1,2]. Recently, there has been increasing evidence that adenosine has tumor-promoting effects [3-5]. Therefore, as a potential tumor marker, sensitive detection of adenosine plays an important role in understanding its role in tumor cell proliferation and further elucidating its function in clinical diagnosis and treatment of cancer. Traditional methods include high-performance liquid chromatography (HPLC)[6,7], capillary electrophoresis[8], and radioimmunoassay[9], but these methods have some limitations, such as t...

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 Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/682C12Q1/6825C12Q2525/205C12Q2563/143C12Q2563/131C12Q2565/607
Inventor 王磊姜玮孙杰威
Owner SHANDONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com