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Method for detecting thrombin by utilizing quantum dot sensitization upconversion nano material

A technology of quantum dot sensitization and thrombin, which is applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., can solve the problem of low signal-to-background ratio, achieve high-sensitivity detection, large absorption coefficient, and improve the effect of detection signal-to-background ratio

Inactive Publication Date: 2019-12-27
HUBEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art, to provide a method for detecting thrombin by using quantum dot sensitized upconversion nanoparticles, to quantitatively detect thrombin in diluted serum, and to solve the problem of using FRET sensor detection. The problem of low background ratio has realized the highly sensitive detection of thrombin in serum, and the detection limit can reach 0.091nM

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  • Method for detecting thrombin by utilizing quantum dot sensitization upconversion nano material
  • Method for detecting thrombin by utilizing quantum dot sensitization upconversion nano material
  • Method for detecting thrombin by utilizing quantum dot sensitization upconversion nano material

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Embodiment 1

[0039] A method for detecting thrombin using quantum dot sensitized upconversion nanomaterials, comprising the following steps:

[0040] (1) Prepare TBA1-UCNPs solution as described above;

[0041] (2) TBA2 and Ag 2 Coupling of Se QDs: 5 μmol Ag 2 Se QDs were added to 1 mL PBS buffer (10 mM, pH=6.8), and after sonication for 5 min, 10 mg EDC·HCl and 5 mg Sulfo-NHS were added thereto, and shaken at room temperature for 30 min. Activated quantum dots were collected by centrifugation, washed twice with PBS buffer (10 mM, pH=7.2) and dispersed in 1 mL of PBS buffer (10 mM, pH=7.2) containing 2 nmol TBA2. Then, incubate at room temperature for 4 h with shaking to obtain the coupling product. Finally, the coupling product was washed three times with PBS buffer (10 mM, pH=7.2), and dispersed in 1 mL Tris buffer (10 mM, pH=7.4), recorded as TBA2-QDs, and stored at 4°C for future use.

[0042] (3) Draw a thrombin detection standard curve: add 0.01mg of TBA1-UCNPs obtained in step (...

Embodiment 2

[0048] A method for detecting thrombin using quantum dot sensitized upconversion nanomaterials, comprising the following steps:

[0049] (1) Prepare TBA1-UCNPs solution as described above;

[0050] (2) TBA2 and Ag 2 Coupling of Se QDs: 5 μmol Ag 2 Se QDs were added to 1 mL PBS buffer (10 mM, pH=6.8), and after sonication for 5 min, 10 mg EDC·HCl and 5 mg Sulfo-NHS were added thereto, and shaken at room temperature for 30 min. The activated quantum dots were collected by centrifugation, washed twice with PBS buffer (10mM, pH=7.2) and then dispersed in 1mL PBS buffer containing 0.5-3nmol TBA2 (0.5, 1, 2, 3, respectively). (10 mM, pH=7.2). Then, incubate at room temperature for 4 h with shaking. Finally, the obtained coupled products were washed three times with PBS buffer (10 mM, pH=7.2), and dispersed in 1 mL Tris buffer (10 mM, pH=7.4), respectively denoted as TBA2-QDs-0.5, TBA2-QDs-1, TBA2-QDs-2, TBA2-QDs-3, stored at 4°C for future use.

[0051] (3) Draw a thrombin dete...

Embodiment 3

[0057] A method for detecting thrombin using quantum dot sensitized upconversion nanomaterials, comprising the following steps:

[0058] (1) Prepare TBA1-UCNPs solution as described above;

[0059] (2) TBA2 and Ag 2 Coupling of Se QDs: 5 μmol Ag 2 Se QDs were added to 1 mL PBS buffer (10 mM, pH=6.8), and after sonication for 5 min, 10 mg EDC·HCl and 5 mg Sulfo-NHS were added thereto, and shaken at room temperature for 30 min. Activated quantum dots were collected by centrifugation, washed twice with PBS buffer (10 mM, pH=7.2) and dispersed in 1 mL of PBS buffer (10 mM, pH=7.2) containing 2 nmol TBA2. Then, incubate at room temperature for 4 h with shaking, and finally, wash the coupled product three times with PBS buffer (10 mM, pH=7.2), disperse in 1 mL Tris buffer (10 mM, pH=7.4), and store at 4°C for future use .

[0060] (3) Draw a thrombin detection standard curve: add 0.01mg of TBA1-UCNPs obtained in step (1) and 0.016nmol of TBA2-QDs obtained in step (2) to 0.2mL HE...

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Abstract

The invention discloses a method for detecting thrombin by utilizing a quantum dot sensitization upconversion nano material. The method comprises the following steps: (1) preparing upconversion nanoparticles modified by an aptamer TBA1 (UCNPs-TBA1); (2) preparing Ag2Se quantum dots modified by an aptamer TBA2 (Ag2Se QDs-TBA2); (3) drawing a standard curve of thrombin detection; and (4) detecting athrombin concentration in a to-be-detected sample. The method disclosed by the invention solves the problem that signal-to-background ratio is low when an FRET sensor is utilized for detection, high-sensitivity detection on thrombin in the to-be-detected sample is realized, and a detection limit can reach 0.091 nM.

Description

technical field [0001] The invention belongs to the field of biological sensing and analysis, and in particular relates to a fluorescent detection method for thrombin by using quantum dot sensitized up-conversion nanomaterials. Background technique [0002] Rare-earth ion-doped upconversion nanoparticles can continuously absorb two or more low-energy photons and emit one high-energy photon. This property of long-wave excitation and short-wave emission can effectively avoid the interference of autofluorescence and scattered light from biological samples. Therefore, up-conversion nanoparticles are widely used in the field of biological analysis and detection. In addition, due to their ability to avoid simultaneous excitation with other substances, upconversion nanoparticles are often used in conjunction with fluorescence resonance energy transfer (FRET) technology as energy donors for FRET systems, while gold nanoparticles, manganese dioxide, and carbon nanoparticles Nanomate...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/64
Inventor 刘志洪余甜雨
Owner HUBEI UNIV
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