Nanocage probe, application thereof and nucleic acid detection method

A detection method and nanocage technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problem that the pretreatment operation is difficult to meet the sensitivity and accuracy of nucleic acid

Active Publication Date: 2020-10-09
NANJING UNIV OF POSTS & TELECOMM
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the nucleic acid content is low, the large sample requirements and complex pretreatment operations in conventional detection methods are difficult to meet the sensitivity and accuracy of nucleic acid detection.

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
  • Nanocage probe, application thereof and nucleic acid detection method
  • Nanocage probe, application thereof and nucleic acid detection method
  • Nanocage probe, application thereof and nucleic acid detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] This embodiment provides a method for preparing a nanocage probe with nucleic acid recognition function, comprising the following steps:

[0055] Step 1, preparation of silver ball template: dissolve 85mg of polyvinylpyrrolidone (PVP) in 20mL of ultrapure water, stir in a round bottom flask and add 85mg of AgNO 3 Let it dissolve completely. Then 200 μL of sodium chloride solution (5M) was added, and AgCl colloid was obtained after stirring for 15 minutes in the dark. In another clean flask, mix 2.5 mL of 0.5 M NaOH solution and 20 mL of 50 mM ascorbic acid solution, and then drop in 2.5 mL of freshly prepared AgCl colloid. The mixture was stirred for 2 hours in the dark, and the silver spheres were collected by washing by centrifugation.

[0056] Step 2. Synthesis of gold / silver nanocages: Dilute the silver nanospheres prepared according to step 1 into 100ml of water, add 200mg of PVP, and heat to boiling for 10 minutes. Then 1.5 mL of HAuCl was added dropwise 4 sol...

Embodiment 2

[0060] This example is the modification of the surface of nanocage particles and its application in dark field imaging monitoring. This embodiment takes microRNA-21 as an example.

[0061] Step 1. According to Example 1, a nanocage probe with nucleic acid recognition function was prepared, and the plasma probe was dispersed and fixed in a detection cell on a positively charged glass surface, thereby obtaining a detection substrate. Add 20 μL of the solution containing microRNA-21 and H2 to the pool and incubate for 1 h. Then incubate with PBS solution containing H3 and H4. After reacting for 1 h, incubate with PBS solution containing heme (50 μM) and K- (0.1 M). Wherein, after each step of incubation, the reaction pool is washed with PBS buffer; finally, G-quadruplex-heme DNase is formed. Such as figure 1 As shown, after the target strand to be tested is hybridized with the hairpin DNA, the hairpin structure opens to form a straight chain. At this time, H2 in the solution ...

Embodiment 3

[0070] This embodiment is the application of the quantitative detection and analysis process of microRNA-21. The detection of microRNA and the acquisition of dark-field signals were carried out according to Example 2, wherein the concentration of microRNA-21 added in step 1 was 20 fM, and other experimental operations and signal recording processes were the same. Under this microRNA-21 concentration, the concentration of 133 particles was analyzed. In response to the light-off signal, count the waiting time of each particle, and use Gaussian fitting to obtain an average waiting time of about 41s, such as Figure 4 shown. Repeat the above steps, experiment with different concentrations of microRNA-21, draw the curve between different concentrations of microRNA-21 and the average waiting time, and fit the linear relationship between the two, such as Figure 5 shown. from Figure 5 It can be seen that the optimal detection concentration of microRNA-21 is at 1×10 -16 ~1×10 -1...

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
particle diameteraaaaaaaaaa
particle diameteraaaaaaaaaa
Login to view more

Abstract

The invention discloses a nanocage probe, an application thereof and a nucleic acid detection method, the nanocage probe comprises gold / silver nanocage particles, the surfaces of the particles are modified with hairpin DNA chains, and the hairpin DNA chains are partially complementary with nucleic acid to be detected; gold / silver nano cage particles are used as a nucleic acid recognition reactionsubstrate and a scattering signal indication probe; a hairpin chain is modified on the surface of the probe as a nucleic acid recognition unit; DNA circulation and HCR amplification processes are realized by utilizing DNA base complementary pairing and strand substitution reaction rules, so that G-quadruplex-heme DNA enzyme is formed on the surface of the probe, etching of a silver component in the nanocage is realized by virtue of active oxygen generated by enzymatic catalysis of hydrogen peroxide decomposition, and a light-off response of a dark field signal is caused. The dark field signalchange with the time resolution capability shows the dependence on the concentration of the nucleic acid to be detected, and quantitative analysis is carried out in combination with a statistical analysis method.

Description

technical field [0001] The invention relates to a probe and its application and a nucleic acid detection method, in particular to a nanocage probe, its application and a nucleic acid detection method. Background technique [0002] As one of the basic substances of life, nucleic acid carries important genetic information and strictly controls protein synthesis and gene expression. The principle of complementary base pairing followed by nucleic acid in the process of recognition and hybridization ensures the strong specificity of nucleic acid detection. Nucleic acid detection has important significance. However, when the nucleic acid content is low, the large sample requirements and complex pretreatment operations in conventional detection methods are difficult to meet the sensitivity and accuracy of nucleic acid detection. Therefore, it is necessary to develop sensitive and accurate nucleic acid detection methods to further shorten the detection window period. Contents of ...

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): C12Q1/6816C12N15/11
CPCC12Q1/6816C12Q2525/301C12Q2563/137C12Q2563/155C12Q2525/207C12Q2563/125C12Q2565/628
Inventor 李美星程娟沈清明范曲立
Owner NANJING UNIV OF POSTS & TELECOMM
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