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Method and enzyme linked immunosorbent assay kit for detecting aflatoxin M1

An enzyme-linked immunosorbent reagent and aflatoxin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that cannot meet the requirements of on-site detection, the detection cost of aflatoxin M1 is expensive, and cannot be quickly detected in large quantities, reaching Improve labeling efficiency, good coating effect, improve precision and stability

Inactive Publication Date: 2015-04-29
TIANJIN BOKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the technical problems that the current detection cost of aflatoxin M1 is relatively expensive, and cannot be quickly detected in large quantities, and cannot meet the requirements of on-the-spot detection.

Method used

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  • Method and enzyme linked immunosorbent assay kit for detecting aflatoxin M1
  • Method and enzyme linked immunosorbent assay kit for detecting aflatoxin M1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, preparation and ELISA detection method using aflatoxin M1 hapten as the coating original kit

[0033] 1. Detection principle:

[0034] First, the aflatoxin M1 antigen is coated on a solid phase carrier, such as a microtiter plate, and then the standard or the sample to be tested is added, and then the enzyme-labeled aflatoxin M1 antibody is added, and the antigen is coated with the standard / sample to be tested Aflatoxin M1 competes with the enzyme-labeled antibody. When the content of aflatoxin M1 in the standard product / sample to be tested is high, the enzyme-labeled antibody that binds to the solid-phase antigen is less, and vice versa. After the reaction, add the substrate solution for color development. When the amount of enzyme-labeled antibody is constant, the more aflatoxin M1 is added in the sample to be tested, the less the enzyme-labeled antibody will bind to the solid-phase antigen, and the color reaction will be weakened. , the percentage abso...

Embodiment 2

[0044] Example 2, the preparation and ELISA detection method of the kit using the aflatoxin M1 specific antibody as the coating source and the antigen as the enzyme marker

[0045] 1. ELISA detection principle using aflatoxin M1-specific antibody as coating source and antigen as enzyme marker:

[0046] When the coating on the microwell strip of the enzyme plate was originally aflatoxin M1-specific antibody, after adding the standard solution or sample solution, and enzyme markers to the microwells of the enzyme plate, the aflatoxin M1 in the sample Compete with the enzyme marker to bind to the specific antibody of aflatoxin M1 on the microwell plate, wash the plate, develop color, the absorbance value of the sample is negatively correlated with the content of aflatoxin M1 in the sample or standard, and can be obtained by comparing with the standard curve The content of aflatoxin M1 in the sample was obtained. At the same time, the content of aflatoxin M1 in the sample can be ...

Embodiment 3

[0110] Embodiment 3, test kit precision, accuracy, preservation test

[0111] 1. The precision test of the kit

[0112] From 3 batches of ELISA plates prepared according to the method in Example 4 (6), 10 microwells were extracted respectively, and the absorbance value (OD value) of the 9 μg / mL standard solution was measured, repeated 10 times, and the coefficient of variation CV was calculated. , the results are shown in Table 1.

[0113] Table 1 Standard solution repeatability test

[0114] cv%

1

2

3

4

5

6

7

8

9

10

01 batch

8

6.5

7.3

6.6

6.9

7.1

8.2

6.8

7.4

8.1

02 batches

8.2

6.8

6.7

6.9

7.2

7.4

8.4

6.3

6.6

8.3

03 batches

7.2

6.8

6.3

6.1

8

7.5

6.3

7.6

8.1

7.7

[0115] The results showed that the intra-assay coefficient of variation of the kit standard product was 6.1-8.4%, and the inter-assay coefficient of variation w...

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Abstract

The invention discloses a method and an enzyme linked immunosorbent assay kit for detecting the content of aflatoxin M1 in a sample. The operation steps can be decreased by adopting direct competitive ELISA detection mode through adopting high-specificity and high-affinity antibodies, and the detection sensitivity and accuracy can be enhanced; compared with the antibody coating, the coating of an ELISA plate is carried out by adopting coating antigen, the better coating effect and long storage time can be more favorably achieved, and further the detection precision and stability of the kit can be enhanced; in addition, according to the kit, marking is carried out by adopting ELISA plate labelled antibody technology through adopting an improved periodate oxidization method, enzyme is directly labelled on an aflatoxin M1 specificity antibody, the two most important reactants namely aflatoxin M1 specificity antibody and enzyme are combined into a whole, so that the labelling efficiency is improved, the usage amounts of the enzyme and the antibody are saved, the good activities of the labelled enzyme and the antibody are guaranteed, the antibody is unnecessarily arranged in the kit, and the cost of the kit is greatly lowered.

Description

technical field [0001] The invention relates to a method for detecting aflatoxin M1 and an ELISA kit. Background technique [0002] Aflatoxins are toxic metabolites of a class of fungi (such as Aspergillus flavus and Aspergillus parasiticus). They are highly carcinogenic and mainly exist in grains, nuts, cottonseed and some products related to human blood and animal feed. Aflatoxin M1 is a hydroxylated metabolite of aflatoxin B1 and is also a strong carcinogen. Milk and its products are one of the foods that are susceptible to contamination by aflatoxin M1. The detection methods of Aflatoxin M1 include high performance liquid chromatography (HPLC), thin layer chromatography (TLC) and so on. The use of aflatoxin M1 ELISA kit can quickly and accurately analyze the residue of aflatoxin M1 in the sample. At present, the methods adopted by the national standard of aflatoxin M1 are thin layer chromatography, spectrophotometer, fluorescence spectroscopy and high performance liqu...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577
CPCG01N33/543G01N33/577
Inventor 王飞
Owner TIANJIN BOKE BIOTECH
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