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Electrochemical immunodetection method of total homocysteine

A technology of homocysteine ​​and immunological detection methods, which is applied in the field of electrochemical analysis technology and immunology, can solve problems such as electrochemical immunological sensing methods that have not yet been seen, achieve the purpose of inhibiting electron transfer, improving analysis and detection performance, and good Dispersion effect

Pending Publication Date: 2022-06-03
SHANGHAI UNIV OF MEDICINE & HEALTH SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing homocysteine ​​detection technology, there is no report on the electrochemical immunosensing method

Method used

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  • Electrochemical immunodetection method of total homocysteine
  • Electrochemical immunodetection method of total homocysteine
  • Electrochemical immunodetection method of total homocysteine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Dissolve 0.04mol TEOS in 200mL absolute ethanol, stir for 1 hour to form a homogeneous solution, add 0.12molNH 3 ·H 2 O and 3.4mol water were stirred overnight at room temperature, centrifuged, washed and dried to obtain SiO 2 Nanospheres. The SiO obtained above was 2 Nanospheres were ultrasonically dispersed in water to obtain a concentration of 10 mg / mL SiO 2 Dispersion, in the ultrasonic process, the above SiO 2 The dispersion was added dropwise to an aqueous PDDA solution with an equal volume concentration of 10 mg / mL, stirred overnight, and centrifuged to obtain positively charged SiO 2 Nanocomposites (PDDA / SiO 2 ). figure 2 where a is the prepared PDDA / SiO 2 SEM picture of nanomaterials, PDDA / SiO can be seen from the picture 2 The surface of the nanomaterials is rough and spherical, and the size is uniform about 150 nm.

[0085] 0.5 mg / mL graphene aqueous dispersion was added dropwise to an equal volume of 4 mg / mL positively charged SiO during sonication...

Embodiment 2

[0105] PDDA / SiO 2 The preparation of nanomaterials is the same as in Example 1, and 1 mL of 1 mg / mL PDDA / SiO 2 The nanomaterial aqueous dispersion was added dropwise to 5 mL of colloidal gold solution with a concentration of 0.079 mg / mL, ultrasonically assembled, centrifuged, and dried to obtain Au-NPs / SiO 2 Nanocomposites.

[0106] The method of constructing the electrochemical immunosensor electrode and using the constructed electrochemical immunosensor to detect homocysteine ​​is the same as that in Example 1.

[0107] Figure 4 The differential pulse curve of the electrochemical immunosensor prepared in this example to SAH standard samples with different concentrations, it can be found by observing the graph that the current response values ​​of the electrochemical immunosensor to 10 μM and 50 μM SAH standard solutions are 22.00 μA and 37.76 μM, respectively. μA. It shows that the nanomaterial prepared in this example can be used to construct an electrochemical immunos...

Embodiment 3

[0109] 0.5g graphene was ultrasonically dispersed in 10mL absolute ethanol, 200μL ammonia water and 0.3g 3-aminopropylethoxysilane were added, refluxed at 75°C for 24h, centrifuged, washed, and dried to obtain amino-functionalized RGO.

[0110] The amino-functionalized RGO was ultrasonically dispersed in water at a concentration of 1 mg / mL, and 1 mL of the above amino-functionalized RGO aqueous dispersion was added dropwise to 5 mL of a colloidal gold solution with a concentration of 0.079 mg / mL, ultrasonically assembled, and centrifuged. The Au-NPs / RGO nanocomposites were obtained by drying.

[0111] The method of constructing the electrochemical immunosensor electrode and using the constructed electrochemical immunosensor to detect homocysteine ​​is the same as that in Example 1.

[0112] Figure 5 The differential pulse curve of the electrochemical immunosensor prepared in this example to SAH standard samples with different concentrations, it can be found by observing the ...

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Abstract

The invention discloses an electrochemical immunodetection method of total homocysteine. The electrochemical immunodetection method comprises the following three steps: preparing an SAH electrochemical immunosensor, drawing a working curve and detecting the total homocysteine. The detection method comprises the following steps: by taking a nano composite material as a raw material, sequentially modifying by streptavidin and a biotinylated SAH antibody and sealing by a bovine serum albumin solution to prepare an electrochemical immunosensor; the electrochemical immunosensor is used for detecting homocysteine in a to-be-detected sample on the basis of the principle that specific reaction of the SAH antibody and the SAH antigen causes change of an electric signal, the total content of homocysteine in the sample is finally obtained, and the electrochemical immunosensor has the advantages of being short in detection time, higher in sensitivity, accuracy and specificity and the like; the rapid, accurate, high-sensitivity, simple and cheap tHcy detection is realized, large-scale detection equipment is not needed, and small-scale intelligent and bedside rapid and simple detection is expected to be realized.

Description

technical field [0001] The invention relates to the technical field of electrochemical analysis technology and immunology, in particular to an electrochemical immunodetection method for total homocysteine. Background technique [0002] Homocysteine ​​(Hcy) is a naturally occurring sulfur-containing amino acid, an important intermediate in the metabolic process of methionine and cysteine, partly as homocysteine-cysteine ​​disulfide. Exist, a small amount exists as reduced homocysteine, and most of it exists through the combination of disulfide bonds with albumin. A large number of studies on Hcy have shown that Hcy is an independent risk factor for cardiovascular and cerebrovascular diseases such as atherosclerosis and thrombosis, as well as vascular thrombosis diseases. Abnormal factors such as genetics, diet and nutrition can cause excessive accumulation of Hcy, which greatly increases the incidence of some diseases such as homocystinuria, Down syndrome, Alzheimer's diseas...

Claims

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Application Information

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IPC IPC(8): G01N27/30G01N27/327G01N33/68
CPCG01N27/308G01N27/3278G01N27/3277G01N33/6815
Inventor 贾冬玲张宜凡赵芳刘敏敏
Owner SHANGHAI UNIV OF MEDICINE & HEALTH SCI
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