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Preparation method and application of molecular imprinting ratio type fluorescence sensor for detecting hepatitis B virus

A hepatitis B virus and fluorescence sensor technology, applied in the field of analytical chemical detection, can solve the problems of limiting the imprinting efficiency of macromolecular viruses, and achieve the effects of low detection limit, high specific surface area, and high specific recognition ability

Active Publication Date: 2020-09-18
XIANGTAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional molecular imprinting techniques mostly use Fe 3 o 4 1. Quantum dots are imprinted carriers. Although they have sufficient magnetic properties or photostability, their relatively small specific surface area will limit the imprinting efficiency of macromolecular viruses.

Method used

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  • Preparation method and application of molecular imprinting ratio type fluorescence sensor for detecting hepatitis B virus
  • Preparation method and application of molecular imprinting ratio type fluorescence sensor for detecting hepatitis B virus
  • Preparation method and application of molecular imprinting ratio type fluorescence sensor for detecting hepatitis B virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: A method for preparing a molecularly imprinted ratiometric fluorescent sensor for dual recognition and detection of HBV.

[0030] (1) MIL-101-NH 2 Preparation: 800mg (2mmol) Cr(NO) 3· 9H 2 O, 360 mg (2 mmol) NH 2 BDC and 159.02 mg NaOH were added into 15 mL water and stirred for 1 hour, then the solution was transferred to a Teflon-based reactor and reacted at 150 °C for 12 hours. After the reaction, the reactor was taken out and naturally cooled to room temperature to obtain a green suspension.

[0031] (2)MIL-101-NH 2 Purification: first wash the obtained MIL-101-NH with DMF solvent 2 Crude product, most of the unreacted impurities were removed. Then solvent heat treatment was used to further wash away a small amount of impurities in the imprinted wells: the product was dispersed in 25mL of ethanol and placed in an oven, reacted at 90°C for 6 hours and then taken out to obtain pure MIL-101-NH 2 .

[0032] (3) Preparation of MIL-101-Apt: first use ED...

Embodiment 2

[0035] Example 2: Morphological and structural characterization of the molecularly imprinted ratiometric fluorescent sensor and intermediate products.

[0036] Using Fourier transform infrared spectrometer, scanning electron microscope, X-ray diffractometer (XRD), thermogravimetric analyzer and N 2 The structure and morphology of all the prepared materials were characterized by adsorption-desorption experiments.

[0037] figure 2 Infrared spectra of MIL-101, MIL-Apt, MIP and NIP. 2976 and 2365cm -1 is the C-H stretching vibration peak of the benzene ring, 1600cm -1 is the amino N-H bending deformation vibration peak, 1498 and 1431cm -1 at MIL-101-NH 2 The stretching and stretching vibration peaks of -(OCO)- in the framework, 1340 and 1258cm -1 is the C-N stretching vibration peak on the aminobenzene ring, indicating that MIL-101-NH 2 The material was prepared successfully, forming NH 2 BDC skeleton. 1600cm -1 The stretching vibration peak of the aptamer C=O is at 12...

Embodiment 3

[0042] Example 3: Application of the molecularly imprinted ratiometric fluorescent sensor.

[0043] The experimental conditions of this example are: the dosage of aptamer is 250 μL, the dosage of monomer TEOS is 40 μL, the concentration of MIP is 0.8 mg / L, the pH is 7.4, the adsorption time is 20 min, and the temperature is 25°C. The specific implementation method is: take a specific concentration of HBV and add it to 0.8 mg / L MIP solution, adjust the pH of the whole system to 7.4, and measure the fluorescence intensity after shaking and absorbing at 25°C for 20 minutes.

[0044] (1) Detection of different concentrations of HBV by MIL-101-MIP ratiometric fluorescent sensor

[0045] According to the above experimental steps, the HBV solutions of different concentrations are detected and analyzed with the ratiometric fluorescent sensor of the present invention, and the results are as follows: Figure 8 As shown, the fluorescence intensity of MIP at 570nm changes, while the fluo...

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Abstract

The invention provides a preparation method of a molecular imprinting ratio type fluorescence sensor for detecting hepatitis B virus. According to the preparation method, a metal organic framework material (MIL-101) is taken as an imprinting carrier, the surface of the imprinting carrier is modified with a virus aptamer, a fluorescent dye monomer rhodamine B and a template virus are added for imprinting, and the molecularly imprinted polymer with aptamer and molecularly imprinted cavity dual recognition is obtained after a template is eluted. After the imprinted polymer is combined with a target object, the fluorescence intensity of a fluorescent dye rhodamine B is changed, and the fluorescence of MIL-101 is almost unchanged, so that a ratio type fluorescence detection system is constructed. According to the method, the advantages of a metal organic framework material, a molecular imprinting technology and fluorescence ratio type detection are combined, the detection sensitivity is high, the detection limit is low, the selectivity is good, the background signal is low, the imprinting factor is as high as 5.72, and the method has a good application prospect in the aspect of virus detection.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry detection, and in particular relates to a preparation method and application of a molecularly imprinted ratiometric fluorescent sensor for detecting hepatitis B virus (HBV). Background technique [0002] Compared with small molecule detection, insufficient selectivity and sensitivity are often encountered in the field of macromolecule detection. Over the years, researchers have developed a variety of strategies to solve this problem and achieved certain results. Including molecular imprint recognition strategy [Liang-hui Luo, Feng Zhang, Chun-yan Chen, Chang-qun Cai.AnalyticalChemistry.2019, 91(24), 15748-15756.], DNA or RNA aptamer recognition strategy [Jae Kwon ,YeonjuLee, Taek Lee, Jae-Hyuk Ahn.Analytical Chemistry.2020,92(7),5524-5531.], compared with traditional antibody recognition strategies, molecular imprinting technology and aptamer technology are both easy to synthesize in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06C08G83/00
CPCG01N21/6486C09K11/06C08G83/008
Inventor 蔡昌群陈思宇罗谅晖陈小明
Owner XIANGTAN UNIV
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