Method for detecting aflatoxin M1

A technology of aflatoxin and detection method, which is applied in the field of antigen detection, can solve the problem of low precision of detection method and achieve the effect of cost saving

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

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Problems solved by technology

[0005] The present invention aims at the technical defect of prior art, provides a kind of anti-afl...

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0054] Detection of Aflatoxin M 1 The preparation and detection method of the new fluorescent ELISA kit, including anti-aflatoxin M 1 Monoclonal Antibody Coated 96-well Black Fluorescent Microtiter Plate, Aflatoxin M 1 Standard, catalase-labeled aflatoxin M 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 M 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 U.S.; anti-aflatoxin M 1 The monoclonal antibody was purchased from Wuxi Zhongdeboer Biotechnology Co., Ltd.; the catalase (cat.No...

Embodiment 2

[0088] Embodiment 2 (aflatoxin M with horseradish peroxidase as antibody labeling enzyme and TMB as chromogenic substrate 1 Direct competition ELISA detection method)

[0089] Traditional ELISA Kit for Detection of Aflatoxin M in Milk Powder and Dairy 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.

[0090] (1) Sample pretreatment

[0091] 1) Dissolve the milk powder first, and then extract with 75% methanol aqueous solution; 2) Shake vigorously for 30 minutes to make it fully mixed; 3) Centrifuge at 5000rpm for 10 minutes, discard the precipitate; 4) Take 1 mL of the supernatant and dilute it with 1 mL of LPBS, and set aside .

[0092] (2) Detect aflatoxin M in the above samples with a traditional ELISA kit 1 residual amount

[0093] anti-aflatoxin M 1 For the microtiter plate of monoclonal antibody, add 50 μL / well of standard / sample to t...

Embodiment 3

[0100] A kind of against aflatoxin M 1 The detection method, this method belongs to the direct competition enzyme-linked immunoassay, this method is for aflatoxin M 1 Antigen detection, the enzyme used to label the antigen in this method is catalase C100.

[0101] On the basis of the above technical solutions, the following conditions are met:

[0102] The substrate in the method includes hydrogen peroxide and mercaptopropionic acid modified cadmium telluride quantum dots.

[0103] The specific detection method includes the following steps:

[0104] 1) coated anti-aflatoxin M 1 Monoclonal antibody, then added to the sample to be tested;

[0105] 2) Then add the aflatoxin M labeled with catalase C100 1 , after mixing, react in a dark environment at 35°C for 40 minutes, and wash;

[0106] 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;

[0107] 4) Then add mercaptopropionic acid-modifi...

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Abstract

The invention provides a method for detecting aflatoxin M1. The method is based on a direct competitive ELISA (enzyme-linked immunosorbent assay) technique, firstly, after monoclonal antibodies are coated, a to-be-detected sample and aflatoxin M1 labeled with catalase C100 are added, aflatoxin M1 in the sample and the aflatoxin M1 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 M1 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, further relates to the antigen detection technology based on ELISA, in particular to a kind of anti-aflatoxin M 1 detection method. Background technique [0002] Aflatoxin is a secondary metabolite produced by fungi such as Aspergillus flavus and Aspergillus parasiticus, mainly aflatoxin B 1 , B 2 , G 1 , G 2 , M 1 and M 2 Wait. Studies have shown that mammalian ingestion of aflatoxin B 1 After contaminated feed or food, aflatoxin B 1 Metabolism in the body produces aflatoxin M 1 . Aflatoxin M 1 It has strong hepatotoxicity, carcinogenicity and mutagenicity to humans and animals. In 2002, the International Agency for Research on Cancer (IARC) classified aflatoxin M 1 Positioned as a class 1 carcinogen. Since it is mainly found in dairy products consumed daily by people, especially infants and young children who are more vulnerable to toxin damage, aflatoxin M 1 Th...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/535G01N33/577
Inventor 熊勇华江湖黄小林裴可熊颖许恒毅
Owner NANCHANG UNIV
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