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Method for detecting aflatoxin M1

An aflatoxin and reaction technology, which can be used in measurement devices, instruments, particle size analysis, etc., can solve the problems of low detection sensitivity, inability to meet detection requirements, and inability to meet the detection requirements of trace aflatoxin M, so as to reduce costs. , the effect of high molar extinction coefficient

Pending Publication Date: 2019-01-25
WUXI ZODOLABS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection kit that this invention provides still can't satisfy trace aflatoxin M 1 testing requirements
[0006] CN104569381A discloses a kind of aflatoxin M in detection sample 1 Content method and enzyme-linked immunoimmunoassay kit, the invention adopts the direct competition ELISA detection mode, and adopts the coated antigen to coat the enzyme-labeled plate, adopts the improved periodate oxidation method to carry out the enzyme-linked immunosorbent plate label, and the enzyme Directly labeled with aflatoxin M 1 Aflatoxin M 1 The two most important reactants of specific antibody and enzyme are combined into one, which improves the labeling efficiency, saves the amount of enzyme and antibody, and ensures that the enzyme and antibody have good activity after labeling, and there is no need to reconfigure the antibody in the kit. The cost of the kit is greatly reduced, but the detection sensitivity is not high, and it cannot meet the requirements of trace aflatoxin M 1 testing requirements

Method used

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  • Method for detecting aflatoxin M1
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  • Method for detecting aflatoxin M1

Examples

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Effect test

Embodiment 1

[0106] The present embodiment provides a method of using glucose oxidase and aflatoxin B 1 Hapten Conjugated as Competing Antigen and Glucose as Substrate for Detection of Aflatoxin M in Milk by Direct Competitive ELISA Based on Dynamic Light Scattering 1 method of residue. The specific operation method is as follows:

[0107] (1) Formal test:

[0108] Dilute protein G to 20 μg / mL with 0.05 mol / L carbonate buffer solution with pH=9.6, and add 100 μL / well of the diluted solution to a 96-well microplate, and let it stand at 4°C for 12 hours; remove the enzyme label The liquid in the plate was washed three times with 0.01mol / L PBS containing 0.05% Tween 20, and then washed once again with 0.01mol / L PBS; 0.01mol / L pH=7.4 Anti-aflatoxin M at a concentration of 0.1 μg / mL diluted in phosphate buffer 1 For monoclonal antibody, let stand at 37°C for 1 hour; remove the liquid in the microtiter plate, wash the microtiter plate three times with 0.01mol / L PBS containing 0.05% Tween 20,...

Embodiment 2

[0120] The present embodiment provides a method of using glucose oxidase and aflatoxin B 1 Hapten Conjugated as Competing Antigen and Glucose as Substrate for Detection of Aflatoxin M in Milk by Direct Competitive ELISA Based on Dynamic Light Scattering 1 method of residue. The specific operation method is as follows:

[0121] (1) Formal test:

[0122]Dilute protein G to 18 μg / mL with 0.04 mol / L carbonate buffer solution with pH=9.4, and add 100 μL / well of the diluted solution to a 96-well microplate, and let it stand at 0°C for 8 hours; remove the enzyme label The liquid in the plate was washed three times with 0.01mol / L PBS containing 0.01% Tween 20, and then washed once again with 0.01mol / L PBS; 0.01mol / L pH=7.4 Anti-aflatoxin M at a concentration of 0.5 μg / mL diluted in phosphate buffer 1 For monoclonal antibody, stand at 35°C for 2 hours; remove the liquid in the microtiter plate, wash the microtiter plate three times with 0.01mol / L PBS containing 0.01% Tween 20, and ...

Embodiment 3

[0134] The present embodiment provides a method of using glucose oxidase and aflatoxin B 1 Hapten Conjugated as Competing Antigen and Glucose as Substrate for Detection of Aflatoxin M in Milk by Direct Competitive ELISA Based on Dynamic Light Scattering 1 method of residue. The specific operation method is as follows:

[0135] (1) Formal test:

[0136] Dilute protein G to 22 μg / mL with 0.06 mol / L carbonate buffer solution of pH=9.8, and add 100 μL / well of the diluted solution to a 96-well microtiter plate, and let it stand at 8°C for 12 hours; remove the enzyme label The liquid in the plate was washed three times with 0.01mol / L PBS containing 0.06% Tween 20, and then washed once again with 0.01mol / L PBS; 0.01mol / L pH=7.4 Anti-aflatoxin M diluted in phosphate buffer at a concentration of 1 μg / mL 1 For monoclonal antibody, stand at 39°C for 3 hours; remove the liquid in the microtiter plate, wash the microtiter plate three times with 0.01mol / L PBS containing 0.06% Tween 20, ...

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Abstract

The invention relates to a method for detecting aflatoxin M1. According to the method, aflatoxin B1 marked by enzyme is taken as a competitive antigen, and on the basis of a dynamic light scattering technology, the aflatoxin M1 is detected through a direct competitive enzyme linked immunoassay analysis method. The method reduces the affinity between the competitive antigen and a monoclonal antibody of the aflatoxin M1, and the problem is solved that the sensitivity is reduced due to the fact that the competitive antigen is not easily competed by s target competitive antigen; the detection of an output signal is performed by means of the dynamic light scattering method, compared with a traditional direct competitive enzyme linked immunoassay analysis method, the detection sensitivity can beimproved, and the trace detection of the aflatoxin M1 can be achieved.

Description

technical field [0001] The invention belongs to the technical field of antigen detection, in particular to a method for detecting aflatoxin M 1 Methods. Background technique [0002] Aflatoxin M 1 Mainly produced by Aspergillus flavus and Aspergillus parasitica, it is one of the structural analogues of aflatoxin. Its toxicity is higher than that of potassium cyanide and arsenic. It is a highly toxic substance and has strong carcinogenicity and mutagenicity. Studies have shown Incidence of liver cancer in high-incidence areas and aflatoxin M in patients' urine 1 closely related. Ingestion of aflatoxin B by mammals 1 After contaminated feed or food, B 1 Convertible to M in animal liver 1 , thereby contaminating the milk. Therefore, monitoring of aflatoxin M in food, especially dairy products 1 has great significance. [0003] due to aflatoxin M 1 Most of the pollution occurs in milk and dairy products, and aflatoxin M 1 Relatively stable, so strict aflatoxin M 1 Li...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/543G01N33/577G01N15/00G01N15/02
CPCG01N15/00G01N15/02G01N15/0211G01N33/535G01N33/54346G01N33/577G01N2015/0222G01N2015/0277G01N2015/0038G01N15/01
Inventor 湛胜楠熊勇华李林袁杰
Owner WUXI ZODOLABS BIOTECH
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