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Method for detecting thrombin based on barometer

A technology of thrombin and barometer, applied in the field of analytical chemistry, can solve the problems of inability to meet the requirements of trace detection, high detection limit, large-scale instrument, etc., achieve good activity and stability, simple method, and compact instrument

Active Publication Date: 2015-11-11
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Thrombin is an important physiological protease that plays an important role in physiological and pathological processes such as blood coagulation, inflammation and wound healing. Striad[J].JThrombHaemost, 2006.4(1):58-59.), immunoassay method (Rand, M.D., Lock, J.B., vantVeer, C., Gaffney, D.P., and Mann, K.G, Blooddottinginimallyalteredwholeblood[J].Blood, 1996.88 (9): 3432—3445.) and fluorescence method (Haixin Chang, Longhua Tang, Ying Wang, Graphene Fluorescence Resonance Energy Transfer Apta sensor for the Thrombin Detection [J]. Anal. Chem. 2010,82,2341–2346), etc., but these methods have high detection limits and cannot meet trace Detection requirements, while the instrument is large and costly, it is difficult to popularize

Method used

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  • Method for detecting thrombin based on barometer
  • Method for detecting thrombin based on barometer
  • Method for detecting thrombin based on barometer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. The establishment of the linear equation of thrombin concentration and pressure with known concentration

[0043] (1) Using catalase (CAT) to modify a section of DNA aptamer A of thrombin to obtain a final concentration of 0.3 μM M CAT-DNA aptamer A solution;

[0044] (2) Add the magnetic beads modified with streptavidin (hereinafter referred to as SA) to the PBS buffer of 0.1 MpH=7.3 containing 0.1M NaCl and 0.05% Tween-20, and then add the magnetic beads modified with biotin (Vh). A section of DNA aptamer B of thrombin was shaken at room temperature for 30 minutes, washed to a constant volume to obtain a DNA aptamer B solution modified on magnetic beads with a final concentration of 0.3 μM;

[0045] (3) Mix the CAT-DNA aptamer A solution prepared in step (1) with the DNA aptamer B solution modified on the magnetic beads in an equal volume ratio, add thrombin containing 5nM (x1) to be tested Solution, incubated at room temperature for 1.5 to 2 hours, due to the spe...

Embodiment 2

[0064] 1. The establishment of the linear equation of thrombin concentration and pressure with known concentration

[0065] With embodiment 1.

[0066] 2. Pretreatment of the thrombin solution sample in the simulated serum solution to be tested and the acquisition of the air pressure value

[0067] Using the same steps as (1) to (6) in the technical scheme "Concentration of Thrombin with Known Concentration and Pressure Linear Equation", the thrombin solution in the serum simulated solution is processed to obtain the corresponding gas pressure value of 0.78KPa (Y );

[0068] 3. Obtaining the concentration (X) of the thrombin solution sample in the serum simulated solution

[0069] According to the linear equation: Y=-0.004+0.0528X, the barometer value Y=0.78KPa of the sample to be tested obtained in "pretreatment of the sample to be tested and obtaining the air pressure value" is substituted into the equation to calculate the thrombin to be tested The concentration of the s...

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Abstract

The invention relates to a method for detecting thrombin based on a barometer, and belongs to the field of analytical chemistry. The method comprises the steps: first modifying catalase on an aptamer of the thrombin, then modifying another section of aptamer B of the thrombin on a magnetic ball, and taking the two sections of aptamers as a capturing recognition probe. When the thrombin exists, a polymer of a sandwiched structure of magnetic ball / aptamer B-thrombin-aptamerA / CAT can be formed, the polymer can be migrated and concentrated under the action of a magnetic field, so that the polymer is convenient to separate and purify; CAT can catalyze a hydrogen peroxide solution to be decomposed to generate hydrogen, thus causing the variation of air pressure in a closed environment, thereby being suitable for the detection application of the barometer. According to the method, the barometer that is low in price is used for substituting a traditional analytical detection tool as a detection means; the thrombin aptamer is used for efficiently recognizing and capturing the specificity of the target thrombin, and a novel method which is low in detection cost, simple to operate, accurate in quantization result and capable of immediately and rapidly detecting the thrombin is established.

Description

technical field [0001] The invention relates to a method for detecting thrombin based on a barometer, belonging to the field of analytical chemistry. technical background [0002] Portable point-of-care detectors generally have the characteristics of small equipment, low detection cost, simple operation, accurate quantitative results, and instant and rapid detection. Compared with traditional analysis and detection methods, portable instant detectors can get rid of the constraints of time, space, equipment cost and operation skills in the detection and analysis process to a certain extent, and can complete low-cost, timely, simple and accurate analysis and detection. It is an analytical technique that does not have too high requirements for experimental testing conditions and operators. It has now attracted people's attention and attention, and has been developed to a certain extent. Common portable instant detectors include thermometers, hygrometers, pressure gauges, infra...

Claims

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Application Information

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IPC IPC(8): G01N33/573
CPCG01N33/573G01N2333/974
Inventor 王清萍李荣杰
Owner FUJIAN NORMAL UNIV
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