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Fabrication method and application of electrochemical immunosensor for detecting brucellosis

An immunosensor and brucellosis technology, which is applied in the field of preparation of electrochemical immunosensors, can solve the problems of poor accuracy of brucellosis detection methods, high requirements for professional background knowledge, and low detection limit of detection range, etc. Achieve the effect of low professional background knowledge requirements, huge commercial value, and wide detection range

Inactive Publication Date: 2019-01-25
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the problems of poor accuracy, complex processing process, and high requirements for the professional background knowledge of operators in the existing brucellosis detection methods, the present invention prepares an electrochemical immunosensor, which can detect Brucella in complex samples Antibody, with ultra-high sensitivity, wide detection range and low detection limit; its preparation method is relatively simple, easy to operate, suitable for mass production

Method used

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  • Fabrication method and application of electrochemical immunosensor for detecting brucellosis
  • Fabrication method and application of electrochemical immunosensor for detecting brucellosis
  • Fabrication method and application of electrochemical immunosensor for detecting brucellosis

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1 Preparation of Electrochemical Immunosensor

[0031] (1) GCEs with a diameter of 3.0 mm were polished into a mirror-like surface with 0.3 μm and 0.05 μm alumina slurries in turn, and then ultrasonically treated in ultrapure water, 100% ethanol and ultrapure water;

[0032] (2) Dip the pretreated electrode into 5mM HAuCl 4 In solution, HAuCl was reduced by potentiostatic 4 , uniformly dispersed gold nanoparticles are generated on the surface of the electrode, the constant potential is -0.2V, and the electrodeposition is applied for 60s;

[0033] (3) The ultrapure water was pre-deoxygenated with nitrogen for 30 min, and then used to prepare the GSH solution to prevent the -SH group from being oxidized. Immerse AuNPs / GCE in the above GSH (400 μl, 10 mM) solution for 48 h, modify GSH to the AuNPs / GCE interface by forming Au-S bonds to obtain GSH / AuNPs / GCE, and then place the modified electrode on β-mercaptoethylamine (200μl, 10mM) for 48h to block unreacted AuN...

Embodiment 2

[0036] Embodiment 2 Preparation of electrochemical immunosensor

[0037] (1) GCEs with a diameter of 3.0 mm were polished into a mirror-like surface with 0.3 μm and 0.05 μm alumina slurries in turn, and then ultrasonically treated in ultrapure water, 100% ethanol and ultrapure water;

[0038] (2) Dip the pretreated electrode into 20mM HAuCl 4 In solution, HAuCl was reduced by potentiostatic 4 , uniformly dispersed gold nanoparticles are generated on the surface of the electrode, the constant potential is -0.2V, and the electrodeposition is applied for 30s;

[0039] (3) The ultrapure water was pre-deoxygenated with nitrogen for 30 min, and then used to prepare the GSH solution to prevent the -SH group from being oxidized. Immerse AuNPs / GCE in the above GSH (400 μl, 10 mM) solution for 52 h, modify GSH to the AuNPs / GCE interface by forming Au-S bonds to obtain GSH / AuNPs / GCE, and then place the modified electrode on β-mercaptoethylamine (200μl, 10mM) for 40h to block unreacted...

Embodiment 3

[0042] Example 3 Preparation of Electrochemical Immunosensor

[0043] (1) GCEs with a diameter of 3.0 mm were polished into a mirror-like surface with 0.3 μm and 0.05 μm alumina slurries in turn, and then ultrasonically treated in ultrapure water, 100% ethanol and ultrapure water;

[0044] (2) Dip the pretreated electrode into 1mM HAuCl 4 In solution, HAuCl was reduced by potentiostatic 4 , uniformly dispersed gold nanoparticles are generated on the surface of the electrode, the constant potential is -0.2V, and the electrodeposition is applied for 300s;

[0045] (3) The ultrapure water was pre-deoxygenated with nitrogen for 30 min, and then used to prepare the GSH solution to prevent the -SH group from being oxidized. Immerse AuNPs / GCE in the above GSH (400 μl, 10 mM) solution for 56 h, modify GSH to the AuNPs / GCE interface by forming Au-S bonds to obtain GSH / AuNPs / GCE, and then place the modified electrode on β-mercaptoethylamine (200μl, 10mM) for 48h to block unreacted Au...

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Abstract

The invention provides a fabrication method of an electrochemical immunosensor for detecting brucellosis, and belongs to the technical field of an electrochemical biosensor. The method comprises the steps of pre-processing a glassy carbon electrode, modifying a gold nanoparticle onto the glassy carbon electrode by a method of electrodeposition, introducing glutathione as a dirt-resistant material,self-assembling with a gold surface, effectively rejoining HA onto the glutathione by N-(3-dimethy inaminoproyl)-N'-ethyl carbodiimide and N-hydroxysuccinimide, and finally, enclosing a non-speical active site by erythritol. By the method, the problems of poor accuracy, complicated processing process and the like of an existing brucellosis detection method are solved. An electrode interface of the immunosensor has excellent hydrophilcity, the pollution-prevention property of a composite interface is obviously improved, the method has the advantages of ultrahigh sensitivity, relatively wide linear range, relatively low detection limit, good stability and repeatability, is simple and convenient to operate and can be used for detecting a brucellosis antibody in a complicated biological sample.

Description

technical field [0001] The invention belongs to the technical field of electrochemical biosensors, and in particular relates to a preparation method and application of an electrochemical immunosensor for detecting brucellosis. Background technique [0002] Brucella is a Gram-negative coccus that causes brucellosis, one of the most common zoonotic diseases. The biggest feature of this disease is that if it is found early and treated with regular methods, more than 90% of patients will be cured and will not relapse throughout their lives, otherwise it will become a chronic disease and cannot be cured throughout their lives. Therefore, early diagnosis of the disease is particularly important. [0003] Typical methods for the detection of brucellosis are tiger red plate agglutination test (RBPT), polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), loop-mediated isothermal amplification technique (LAMP) and immunoassay Chromatography test. Among them, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N27/327
CPCG01N27/308G01N27/3278
Inventor 陈丽华赵师伊李聪年
Owner QINGDAO UNIV OF SCI & TECH
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