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A kind of preparation method of the sandwich type electrochemical immunosensor of cardiac troponin

A technology of cardiac troponin and immunosensor, which is applied in the field of immunoassay and biosensing, can solve the problems of long detection time, complicated operation, radioactive contamination, etc., and achieve the goal of lower detection limit, larger specific surface area, and excellent conductivity Effect

Inactive Publication Date: 2020-08-11
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many detection methods for cardiac troponin, such as enzyme-linked immunosorbent assay, immunofluorescence, etc., but these methods require large-scale equipment during the detection process, the operation is complicated, the detection takes a long time, and there are problems such as radioactive contamination. Therefore, it is very important to invent a cardiac troponin immunosensor with strong specificity, high sensitivity, fast detection speed and easy operation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1 prepares gold nanocubic functionalized graphene oxide dispersion, the steps are as follows:

[0051] (1) Preparation of gold nanocubic dispersion

[0052] Prepare 10 mL of a mixture containing 0.25 mM chloroauric acid and 75 mM cetyltrimethylammonium bromide in a 50 mL round bottom flask, and quickly add 0.5 mL, 10 mM freshly prepared ice boron under magnetic stirring Sodium hydride solution was stirred in a water bath at 27°C for 3 hours to obtain a gold nanoparticle aqueous sol;

[0053] Measure 1.0 mL of the above-prepared gold nanoparticle hydrosol and dilute it to 100 mL with ultrapure water as the gold seed dispersion;

[0054] In a 50 mL centrifuge tube, add 0.1 mL, 10 mM chloroauric acid solution, 2.0 mL, 0.2 M cetyltrimethylammonium bromide solution, 1.5 mL, 0.1 M ascorbic acid, and 0.3 mL of the above-prepared gold seed dispersion solution, diluted to 25 mL with ultrapure water, and shaken at 27 °C for 8 hours in a constant temperature shaking i...

Embodiment 2

[0059] Embodiment 2 prepares gold nanocubic functionalized graphene oxide dispersion, the steps are as follows:

[0060] (1) Preparation of gold nanocubic dispersion

[0061] Prepare 10 mL of a mixture containing 0.25 mM chloroauric acid and 75 mM cetyltrimethylammonium bromide in a 50 mL round bottom flask, and quickly add 0.6 mL of 10 mM freshly prepared ice boron under magnetic stirring Sodium hydride solution was stirred in a water bath at 27°C for 3 hours to obtain a gold nanoparticle aqueous sol;

[0062] Measure 1.0 mL of the above-prepared gold nanoparticle hydrosol and dilute it to 100 mL with ultrapure water as the gold seed dispersion;

[0063] In a 50 mL centrifuge tube, add 0.1 mL, 10 mM chloroauric acid solution, 2.0 mL, 0.2 M cetyltrimethylammonium bromide solution, 1.5 mL, 0.1 M ascorbic acid, and 0.4 mL of the above-prepared gold seed dispersion solution, diluted to 25 mL with ultrapure water, and shaken at 27 °C for 8 hours in a constant temperature shaking...

Embodiment 3

[0068] Example 3 Preparation of gold nanocubic functionalized graphene oxide dispersion, the steps are as follows:

[0069] (1) Preparation of gold nanocubic dispersion

[0070] Prepare 10 mL of a mixture containing 0.25 mM chloroauric acid and 75 mM cetyltrimethylammonium bromide in a 50 mL round bottom flask, and quickly add 0.7 mL, 10 mM freshly prepared ice boron under magnetic stirring Sodium hydride solution was stirred in a water bath at 27°C for 3 hours to obtain a gold nanoparticle aqueous sol;

[0071] Measure 1.0 mL of the above-prepared gold nanoparticle hydrosol and dilute it to 100 mL with ultrapure water as the gold seed dispersion;

[0072] In a 50 mL centrifuge tube, add 0.1 mL, 10 mM chloroauric acid solution, 2.0 mL, 0.2 M cetyltrimethylammonium bromide solution, 1.5 mL, 0.1 M ascorbic acid, and 0.5 mL of the above-prepared gold seed dispersion solution, diluted to 25 mL with ultrapure water, and shaken at 27 °C for 8 hours in a constant temperature shakin...

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Abstract

The invention belongs to the technical field of immune analysis and biosensing, and provides a preparation method of a sandwich type electrochemical immunosensor for cardiac troponin. The present invention uses gold-silver core-shell nano-cubic functionalized nitrogen-sulfur double-doped graphene oxide as a catalytic material, and uses it as a marker after incubation with a detection antibody, and simultaneously uses gold nano-cubic functionalized graphene oxide as an electrode modification material, thereby The detection of cardiac troponin cTnI has been realized, which has high sensitivity, strong specificity and low detection limit, and has important scientific significance and application value for the early detection of cardiac troponin.

Description

technical field [0001] The invention belongs to the technical field of immune analysis and biosensing, and relates to a preparation method of a sandwich-type electrochemical immunosensor for cardiac troponin. Specifically, gold-silver core-shell nano-cubic functionalized nitrogen-sulfur double-doped graphene oxide is used as the catalytic material to label the detection antibody, and gold nano-cubic functionalized graphene oxide is used as the electrode modification material, thereby realizing the detection of cardiac muscle calcium. Sensitive detection of proteins. Background technique [0002] Cardiac troponin is currently considered to be a better marker of myocardial damage, which can be used to diagnose myocardial infarction. The level of cardiac troponin in normal blood is below 0.5 ng / mL. 6-9 hours after myocardial infarction, blood center An increase in troponin levels occurs and persists for several days. Therefore, the realization of sensitive detection of cardia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/30G01N27/48G01N33/68G01N33/532
Inventor 李月云吕慧李新进徐振
Owner SHANDONG UNIV OF TECH
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