Method for detecting aflatoxin B1

A technology of aflatoxin and detection method, which is applied in the field of antigen detection and can solve the problems of low precision of detection method

Active Publication Date: 2016-07-13
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical defect of prior art, provides a kind of anti-aflatoxin B 1 A detection method to address prior art aflatoxin B 1 ELISA detection method has low precision

Method used

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  • Method for detecting aflatoxin B1
  • Method for detecting aflatoxin B1
  • Method for detecting aflatoxin B1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1 (based on aflatoxin B of the present invention 1 A new fluorescent ELISA detection method prepared a kit and used it to detect aflatoxin B in corn and corn products 1 residue)

[0054] Detection of Aflatoxin B 1 The preparation and detection method of the new fluorescent ELISA kit, including anti-aflatoxin B 1 Monoclonal Antibody Coated 96-well Black Fluorescent Microtiter Plate, Aflatoxin B 1 Standard, catalase-labeled aflatoxin B 1 Working solution, substrate solution A hydrogen peroxide, fluorescent substrate solution B mercaptopropionic acid modified cadmium telluride quantum dots and concentrated washing solution.

[0055] Detect aflatoxin B in the present invention in detail below 1 Preparation of the ELISA kit,

[0056] The 96-well black fluorescent microplate plate was purchased from Corning Company in the United States; anti-aflatoxin B 1 The monoclonal antibody was purchased from Wuxi Zhongdeboer Biotechnology Co., Ltd.; the catalase (cat.N...

Embodiment 2

[0089] Embodiment 2 (with horseradish peroxidase as antibody labeling enzyme, with TMB as the aflatoxin B of chromogenic substrate 1 Direct competition ELISA detection method)

[0090] Traditional ELISA Kit for the Detection of Aflatoxin B in Corn and Corn Products 1 For the residual amount, it is implemented through the following steps: sample pretreatment, detection with the kit of the present invention, and analysis of the results.

[0091] (1) Sample pretreatment

[0092] Take the pulverized sample and pass it through a 20-mesh sieve, and mix thoroughly; weigh 5g sample, add 12.5mL extract solution (methanol:water=7:3), shake vigorously for 30min; centrifuge at 5000rpm for 10min, filter with filter paper; take 1mL filtrate for Dilute with 1mL LPBS and set aside.

[0093] (2) Detect aflatoxin B in the above samples with a traditional ELISA kit 1 residual amount

[0094] anti-aflatoxin B 1 For the microtiter plate of monoclonal antibody, add 50 μL / well of standard / samp...

Embodiment 3

[0101] aflatoxin B 1 The detection method, which belongs to the direct competition enzyme-linked immunosorbent assay, the method is for aflatoxin B 1 Antigen detection, the enzyme used to label the antigen in this method is catalase C100.

[0102] On the basis of the above technical solutions, the following conditions are met:

[0103] The substrate in the method includes hydrogen peroxide and mercaptopropionic acid modified cadmium telluride quantum dots.

[0104] The specific detection method includes the following steps:

[0105] 1) Coating anti-aflatoxin B 1 Monoclonal antibody, then added to the sample to be tested;

[0106] 2) Then add catalase C100 labeled aflatoxin B 1 , after mixing, react in a dark environment at 35°C for 40 minutes, and wash;

[0107] 3) Then add hydrogen peroxide solution with a concentration of 8 μmol / L to mix, and react for 20 minutes at 35°C in a dark environment;

[0108] 4) Then add mercaptopropionic acid-modified cadmium telluride quan...

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Abstract

The invention provides a method for detecting aflatoxin B1. The method is based on a direct competitive ELISA (enzyme-linked immunosorbent assay) technique, firstly, monoclonal antibodies are coated, a to-be-detected sample and aflatoxin B1 labeled with catalase C100 are added, aflatoxin B1 in the sample and the aflatoxin B1 labeled with catalase C100 are competitively combined with the monoclonal antibodies fixed on an ELISA plate, fluorescence quenching of cadmium telluride quantum dots modified with mercaptopropionic acid is reduced through decomposition of hydrogen peroxide under the catalytic function of catalase, and the content of aflatoxin B1 in the sample is judged according to the fluorescence intensity. According to the method, new catalase is introduced innovatively, and the reaction precision is improved while the cost is reduced; meanwhile, a more sensitive novel fluorogenic substrate, namely, the cadmium telluride quantum dots modified with mercaptopropionic acid, is adopted, and the lighting sensitivity is remarkably improved by comparison with traditional TMB (tetramethylbenzidine) substrates.

Description

technical field [0001] The present invention relates to the technical field of antigen detection, and further relates to ELISA-based antigen detection technology, in particular to a kind of anti-aflatoxin B 1 detection method. Background technique [0002] Aflatoxin is a class of derivatives containing a dihydrofuran ring and an oxone coumarin structure, and is a secondary metabolite produced by fungi such as Aspergillus flavus and Aspergillus parasiticus. Aflatoxin B 1 , B 2 , G 1 , G 2 , M 1 and M 2 etc., of which aflatoxin B 1 It is the most widely distributed in food and is the most harmful to human health. Studies have shown that aflatoxin B 1 It is highly hepatotoxic, carcinogenic, mutagenic and immunosuppressive to humans and animals. International Agency for Research on Cancer (IARC) aflatoxin B 1 Positioned as the most toxic class 1 carcinogen. Aflatoxin B 1 It often contaminates foods such as corn, peanuts, rice, nuts and vegetable oils. Therefore, es...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/535G01N33/577
Inventor 熊勇华江湖黄小林段宏郑玲燕许恒毅
Owner NANCHANG UNIV
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