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Magnetic mesoporous silica nanoparticle probe and its preparation method and application

A technology of mesoporous silica and nano-particles, which is applied in the direction of material analysis and instrumentation by chemical reaction of materials, material analysis by observing the influence of chemical indicators, etc., can solve the complex operation process, operation process error, High operation requirements, to achieve the effect of simple process, wide application field and good stability

Active Publication Date: 2020-09-29
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most of the methods for clinical detection of viral diseases are relatively complicated, and laboratory detection has the disadvantages of high operational requirements, professional operation, time-consuming, and expensive instruments.
The most typical test for the signal amplification effect often used in laboratory testing is the enzyme-linked immunosorbent assay, which can use chemical means to achieve signal amplification, but the operation process of the enzyme-linked immunosorbent assay is complicated and requires professional operation, and the operation process is extremely easy The error caused the deviation of the result

Method used

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  • Magnetic mesoporous silica nanoparticle probe and its preparation method and application
  • Magnetic mesoporous silica nanoparticle probe and its preparation method and application
  • Magnetic mesoporous silica nanoparticle probe and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Taking the detection probe of Newcastle disease virus as an example, constructing a magnetic mesoporous silica nanoparticle probe

[0035] 1. Characterization of M-MSN nanoparticles: Dilute M-MSN nanoparticles to 1μg / μL, then take 10μL spotting copper grid, and then observe the size and pore diameter of nanoparticles with transmission electron microscope and scanning electron microscope. The results are shown in figure 1 and figure 2 .

[0036]2. Loading of peroxidase (POD): the commercialized peroxidase and the diluted M-MSN nanoparticles in step 1 were in a mass ratio of 1:1 (i.e. 100 μg of M-MSN:100 μg of POD ) mixed, and the mixed solution was incubated on the mixture at 4°C for 8-12h; then the unbound peroxidase was separated and washed with a magnetic frame to obtain the supernatant and the precipitate, and the obtained precipitate was POD / M-MSN; the precipitate was dissolved in 100μL 20 μg of M-MSN nanoparticles were dissolved in 100 μL of PBS, and 2...

Embodiment 2

[0041] The detection of embodiment 2 peroxidase activity

[0042] In order to detect the activity of peroxidase in magnetic mesoporous silica, we chose to use TEM as the substrate to react with peroxidase in different states to develop color, and the specific grouping is pure peroxidase (POD ), magnetic mesoporous silica-coated peroxidase (POD / M-MSN), polyethyleneimine-coated magnetic mesoporous silica loaded with peroxidase (POD / M-MSN / PEI) and The completed nanoprobe (POD / M-MSN / PEI / Ab) was constructed, and its OD450 value was read using a microplate reader. The test results found that after the peroxidase was loaded into the magnetic mesoporous silica, there was no change in the enzymatic activity of the peroxidase in each state. The specific test results are as follows:

[0043] POD content OD450 POD0.2μg 3.9267 POD / M-MSN 0.2μg 3.9389 POD / M-MSN / PEI 0.2μg 3.9165 POD / M-MSN / PEI / Ab 0.2μg 3.7982

Embodiment 3

[0044] Example 3 Magnetic Mesoporous Silica Nanoparticle Probe Detection of Newcastle Disease Virus

[0045] 1. The magnetic mesoporous silica nanoparticle probe (POD / M-MSN / PEI / Ab) prepared in Example 1 is combined with Newcastle disease virus: the magnetic mesoporous silica nanoparticle probe is combined with Newcastle disease virus The ratio is 1:1, and react at room temperature for 1 hour. After 1 hour, use a magnetic stand to separate the supernatant and precipitate, and wash 3 times with 1×PBS. Set aside for precipitation.

[0046] 2. Extraction of viral RNA and reverse transcription: extract RNA from the precipitated product obtained in step 1 using an RNA extraction kit, and then use a reverse transcription kit to reverse transcribe the extracted RNA into cDNA. The reverse transcription system is as follows:

[0047]

[0048] The reverse transcription process is 37°C, 15min, 85°C, 5s.

[0049] 3. Detection of binding efficiency: the reverse transcription product i...

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Abstract

The invention discloses a magnetic mesoporous silica nanoparticle probe. The magnetic mesoporous silica nanoparticle probe firstly loads the magnetic mesoporous silica nanoparticle with peroxidase to obtain M-MSN / POD, Then polyethyleneimine is wrapped with M‑MSN / POD to encapsulate peroxidase to obtain M‑MSN / POD / PEI nanoparticles, which are then mixed with antibodies to form magnetic mesoporous silica nanoparticle probes. The invention also discloses a preparation method of the magnetic mesoporous silicon dioxide nano particle probe. The probe can be used for the specific detection of cell targeting, the visual detection of viruses, and the preparation of ultrasensitive detection test strips. Compared with the prior art, the product prepared by the present invention has high specificity and good stability , a wide range of application fields, etc., combined with simple process and low cost, it has a good prospect for promotion.

Description

technical field [0001] The invention belongs to the technical field of preparation of mesoporous silicon dioxide probes, and in particular relates to magnetic mesoporous silicon dioxide nanoparticle probes and their preparation methods and applications. Background technique [0002] Since Kresge et al first reported a series of new mesoporous molecular sieve materials in 1992, mesoporous silica materials have attracted extensive attention of researchers. Mesoporous silica is an excellent carrier for loading small molecules due to its simple synthesis and low cost, large specific surface area, porous structure with adjustable size and structure, and easy chemical modification of internal and external surfaces. [0003] In the past period of time, a series of studies have been carried out on the application of mesoporous silica materials in adsorption, imaging, drug delivery, and biosensors. Mesoporous silica has porous properties and is widely used to carry drugs, proteins, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/535G01N21/78
CPCG01N21/78G01N33/535G01N33/56983
Inventor 周昕李真真
Owner YANGZHOU UNIV