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Thrombin detection method based on bio-dots and Au NPs fluorescence resonance energy transfer

A technology of fluorescence resonance energy and detection method, which is applied in the field of optical sensing, can solve the problems of no thrombin application and no research, and achieve the effects of reduced fluorescence resonance energy transfer effect, good selectivity, and high sensitivity

Inactive Publication Date: 2016-08-10
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no related research on FRET between Au NPs and bio-dots, nor its application in thrombin detection

Method used

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  • Thrombin detection method based on bio-dots and Au NPs fluorescence resonance energy transfer
  • Thrombin detection method based on bio-dots and Au NPs fluorescence resonance energy transfer
  • Thrombin detection method based on bio-dots and Au NPs fluorescence resonance energy transfer

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Experimental program
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Effect test

Embodiment 1

[0019] (1) Preparation of biological quantum dots: Dissolve the thrombin aptamer (TBA) in ultrapure water, transfer the solution to a polytetrafluoroethylene autoclave, and heat it to 100 °C for 12 h to produce thrombin Aptamer biological quantum dots (TBA-dots).

[0020] (2) Preparation of Au NPs: 2 mL of HAuCl 4 (1% wt) and 48 mL of ultrapure water were mixed evenly, transferred to a conical flask, heated to boiling and stirred vigorously; after the solution boiled, 1 mL of sodium citrate (5% wt) solution was quickly added, and the color of the solution changed from light yellow to slow Slowly turn dark red, continue to stir for 5 min, cool to room temperature, prepare Au NPs solution, and store at 4 °C for later use.

[0021] The morphology of TBA-dots was characterized by transmission electron microscopy, and the results were as follows: figure 1 shown. Depend on figure 1 It can be seen that TBA-dots are spherical and have a uniform particle size distribution, with an ...

Embodiment 2

[0023] Detection of Thrombin Based on Biological Quantum Dots and Au NPs Fluorescence Resonance Energy Transfer

[0024] In phosphate buffer (concentration 10 mM, pH 7.4, containing 50 mM NaCl, 5 mM KCl, 4 mM MgCl 2 ), mixed 80 nM TBA-dots with different concentrations of thrombin, incubated at 37 °C for 1 h, added the mixed solution to 2.3 nM Au NPs solution, and measured the fluorescence intensity of TBA-dots (excitation Voltage 700 V, slit width 10 nm, excitation wavelength 320 nm, wavelength scanning range 350-570 nm).

[0025] Depend on image 3 A shows that Au NPs have good dispersion; after adding TBA-dots to the Au NPs solution, TBA-dots are assembled to the surface of Au NPs through the interaction of Au-N bonds and form a "shell" structure to wrap Au NPs, resulting in Au NPs Gather ( image 3 B) At the same time, since the absorption spectrum of Au NPs partially overlaps with the emission spectrum of TBA-dots, the FRET between TBA-dots and Au NPs reduces the fluor...

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Abstract

Belonging to the technical field of optical sensing, the invention discloses a thrombin detection method based on bio-dots and Au NPs fluorescence resonance energy transfer. A thrombin aptamer is employed as the template to prepare bio-dots, the bio-dots are mixed with a to-be-tested thrombin sample, then the mixed solution is added into an Au NPs solution, at the moment, the fluorescence intensity of the bio-dots is directly correlated with the concentration of thrombin, and the concentration of thrombin is judged according to the fluorescence intensity of bio-dots.

Description

technical field [0001] The invention relates to a thrombin detection method based on biological quantum dots and Au NPs fluorescence resonance energy transfer, belonging to the technical field of optical sensing. Background technique [0002] Thrombin is ubiquitous in the human body and is an endoprotease that functions as a coagulation factor in physiological and pathological processes and can convert fibrinogen into fibrin. Thrombin also has hormone-like properties and plays an important role in thrombus formation and platelet activation. Therefore, thrombin plays an important role in many cardiovascular diseases, with the role of regulating inflammation and tissue repair on the blood vessel wall. Thrombin can also be used as a therapeutic agent and biomarker for the diagnosis of various diseases, such as lung metastases and related coagulation abnormalities. Therefore, the establishment of a simple and sensitive thrombin detection method is of great significance for the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 梁汝萍匡岚邱建丁
Owner NANCHANG UNIV
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