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Preparation method and application of a kind of competitive aflatoxin immunosensor

An aflatoxin and immunosensor technology, which can be used in instruments, scientific instruments, measuring devices, etc., can solve the problems of complex procedures, long detection periods, and many required reagents, and achieve improved detection sensitivity and good biocompatibility. Effect

Active Publication Date: 2015-10-21
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to its long detection cycle, complex procedures and many reagents required, it is far from meeting the requirements of modern detection.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation of the hydroxyapatite-labeled antibody loaded with copper ions according to the present invention, the steps are as follows:

[0034] (1) Mix nano-hydroxyapatite with a particle size of 30nm and copper sulfate at a mass ratio of 1:3, dissolve in 5mL water, shake for 12 hours, and centrifuge;

[0035] (2) Take 5 mg of the obtained nano-hydroxyapatite loaded with copper ions, dissolve it in 5 mL of phosphate buffer solution, add 0.1 ml of aflatoxin antibody solution with a concentration of 1 μg / mL, incubate and shake for 4 hours, and centrifuge;

[0036] (3) Re-add the obtained hydroxyapatite-labeled antibody loaded with copper ions into 5 mL of phosphate buffer solution, and add 1 mg of bovine serum albumin, incubate and shake for 4 hours, and centrifuge;

[0037] (4) Redisperse the precipitate into 5 mL of phosphate buffer solution to obtain the stock solution of the hydroxyapatite-labeled antibody loaded with copper ions.

Embodiment 2

[0039] The preparation of the competitive type aflatoxin immunosensor of the present invention, the steps are as follows

[0040] (1) Add 50μL of an aqueous solution containing 0.05mmol / L silver nitrate, 0.2mmol / L sodium citrate and 0.05mol / L potassium nitrate to the screen-printed electrode, connect to the electrochemical workstation, and perform constant potential deposition at -0.2V, Deposition time is 120s, dries in the air, makes the screen-printed electrode of silver nanoparticle modification;

[0041] (2) Add 10 μL of aflatoxin antibody solution with a concentration of 1 μg / mL dropwise on the screen-printed electrode, incubate for 1 hour, and wash it with phosphate buffer solution;

[0042] (3) Add 10 μL of 1% bovine serum albumin solution dropwise on the screen-printed electrode, incubate for 1 hour, and wash it with phosphate buffer solution;

[0043] (4) Add 10 μL of the mixed solution of aflatoxin and copper ion-loaded hydroxyapatite-labeled antigen dropwise at a v...

Embodiment 3

[0045] A kind of competitive type aflatoxin immunosensor described in any one of embodiment 1~2, is used for the detection of aflatoxin, comprises the following steps:

[0046] (1) Prepare a series of screen-printed electrode immunosensors according to the method described in any one of Examples 1-2 by using different concentrations of aflatoxin;

[0047] (2) Connect the reference electrode, counter electrode and working electrode to the electrochemical workstation, add 50 μL of acetate buffer solution to the electrolytic cell, and then add 5~10 μL of 1% dilute sulfuric acid solution to load copper ions The hydroxyapatite dissolves and releases copper ions;

[0048] (3) Set the potential range to -0.6~0.4V, the deposition potential to -0.8V, and the deposition time to 100~300s, and detect the current response of the modified screen-printed electrode by anodic stripping linear voltammetry;

[0049] (4) Draw a working curve according to the relationship between the obtained cur...

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Abstract

The invention belongs to the technical field of food detection and bio-sensing, and discloses a competitive immunosensor for rapidly detecting aflatoxin. A manufacturing scheme and a detection method comprise the following steps: by adopting a screen-printed electrode as a working electrode, modifying a layer of silver nano particles by virtue of an electro-deposition process, then sequentially adding an aflatoxin antibody, bovine serum albumin, aflatoxin and a copper ion supported hydroxyapatite labeled antigen mixed solution, and dissolving copper ion supported hydroxyapatite by using dilute sulfuric acid to ensure that copper ions in copper ion supported hydroxyapatite are released. Therefore, the detection of aflatoxin is transformed into the detection of the copper ions, relatively high sensitivity can be achieved, and the limit of detection can be as low as 0.1ng / kg.

Description

technical field [0001] The invention belongs to the technical field of food detection and biosensing, and in particular relates to a screen printing electrode immunosensor for rapid detection of aflatoxin. Background technique [0002] Aflatoxins are metabolites of Aspergillus flavus and Aspergillus parasiticus, mainly including B1, B2, G1, G2 and two other metabolites M1 and M2. Among them, M1 and M2 are separated from milk. B1, B2, G1, G2, M1 and M2 are very close in molecular structure. [0003] Aflatoxin is the strongest carcinogen found so far, and its carcinogenicity is 75 times greater than the ability of dimethylnitrosamine to induce liver cancer. The half-lethal dose of aflatoxin Bl is 0.36mg / kg body weight, which is extremely toxic range of poisons. It causes human poisoning mainly to damage the liver, hepatitis, liver cirrhosis, and liver necrosis. Clinical manifestations include stomach discomfort, loss of appetite, nausea and vomiting, abdominal distension a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N27/48
CPCG01N33/56961
Inventor 王欢魏琴张以河庞雪辉吴丹张勇马洪敏范大伟
Owner UNIV OF JINAN