Smart hydrogel-based fluorescent aptasensor for detection of avian influenza virus

A technology of aptamer sensor and avian influenza virus, which is applied in the field of fluorescent aptamer sensor and its preparation, can solve the problems of limited field application, QCM is easily affected by environmental factors, etc., and achieves easy modification and immobilization, high Specific, low-cost effects

Active Publication Date: 2017-08-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, Wang and Li immobilized the gel on the surface of QCM sensor to detect avian influenza virus, but the field application of QCM was limited due to the susceptibility of QCM to environmental factors.

Method used

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  • Smart hydrogel-based fluorescent aptasensor for detection of avian influenza virus
  • Smart hydrogel-based fluorescent aptasensor for detection of avian influenza virus
  • Smart hydrogel-based fluorescent aptasensor for detection of avian influenza virus

Examples

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Effect test

Embodiment 1

[0070] Embodiment 1, a kind of preparation method of the fluorescent aptamer sensor of rapid detection avian influenza virus H5N1 based on intelligent response hydrogel, specifically as follows:

[0071] 1. Preparation of single-stranded DNA 2 -Quantum dot coupler

[0072] ssDNA 2 (80 μL, 5 μM) and quantum dot solution (16 μL, 1 μM) were mixed in a small centrifuge tube, incubated at room temperature for 30 min under agitation, and stored at 4°C for use.

[0073] The ssDNA 2 The sequence is: 5'- / 5Biosg / CAA CAG GAC AAC-3'.

[0074] The formula of the quantum dot solution is: cadmium selenide / zinc sulfide CdSe / ZnS core-shell semiconductor fluorescent quantum dots, the solvent is borate buffer solution (0.05M, pH 7.4), and the maximum emission wavelength is 626nm.

[0075] 2. Preparation of single-stranded DNA 1 - Aptamer polymer hydrogels:

[0076] (1), configure 0.2% (mass %) acrylamide aqueous solution as stock solution;

[0077] (2), configure initiator-catalyst mixed ...

Embodiment 2

[0101] Embodiment 2, using the above-mentioned intelligent response hydrogel-based fluorescent aptasensor for rapid detection of different concentrations of avian influenza virus H5N1:

[0102] A portable fluorescence spectrum detection system (referenced from U.S. Patent US 2008 / 0135490 A1) used in conjunction with the aptamer sensor described in the present invention, its composition includes:

[0103] (1) A USB2000 optical detector that can detect wavelengths from 360 to 900nm;

[0104] (2) A portable light source that can emit 380nm excitation light;

[0105] (3) An optical fiber and probe capable of transmitting UV-visible reflected / scattered light;

[0106] (4) a self-made black box that can hold a cuvette;

[0107] (5) And a set of software and laptop computer that can read test data and display test results.

[0108] Its use method is as follows (taking hydrogel A as an example):

[0109] Such as figure 2 As shown, a fluorescent aptasensor based on an intelligent...

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Abstract

The invention discloses a fluorescent aptamer sensor for avian influenza virus detection based on smart hydrogel, and particularly discloses the fluorescent aptamer sensor for quickly detecting the avian influenza virus H5N1 based on smart response hydrogel. According to the method, the DNA nucleic acid aptamer which has a high affinity for the avian influenza virus H5N1 is selected to be used as a biological recognition component, single-stranded DNA 1 and single-stranded DNA 2 which are complementary with the nucleic acid aptamer are designed, DNA chains modified by acrydite can be polymerized to form a polyacrylamide chain containing a plurality of DNA branched chains under the catalysis action of an initiator, and base complementary pairing between the nucleic acid aptamer and the single-stranded DNA 1 plays the effect of a cross-linking agent; when no target object exists, the gel is in a contraction state due to the hybridization action between the DNA chains, and the fluorescence of quantum dots is quenched for the reason that the quantum dots get close to a quenching agent; when the target object exists, the aptamer is combined with the target object, so that hybridization between the DNA chains is decomposed, the gel is swelled, the distance between the quantum dots and the quenching agent is increased, and accordingly the fluorescence of the quantum dots is recovered.

Description

technical field [0001] The invention relates to a method for rapidly detecting viruses, in particular to a fluorescent aptamer sensor for rapidly detecting avian influenza virus H5N1 based on an intelligent response hydrogel and a preparation method thereof. Background technique [0002] Avian influenza is an infectious disease of poultry caused by type A influenza virus. Avian influenza virus is pleomorphic, with a spherical diameter of 80-120nm and a capsule. According to the different antigenicity of outer membrane hemagglutinin H and neuraminidase protein N, it can be divided into 16 H subtypes and 9 N subtypes, among which the subtypes of avian influenza virus infecting humans are mainly H5N1, H9N2, H7N9 etc. The highly pathogenic avian influenza virus H5N1 has threatened global public health security and caused huge economic losses. Since 2003, a total of 844 people from 62 countries / regions around the world have been infected and 449 people have died from it (WHO, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/11
Inventor 李延斌徐李舟王蓉晖
Owner ZHEJIANG UNIV
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