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Aflatoxin enzyme linked immunosorbent assay kit and detecting method

An enzyme-linked immunosorbent reagent and aflatoxin technology, applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of inability to adapt to large sample detection, detection process and operation process troubles, etc.

Inactive Publication Date: 2019-05-07
CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the detection methods of aflatoxin on the market mainly include thin-layer chromatography, high performance liquid chromatography, liquid chromatography-mass spectrometry, enzyme-linked immunosorbent assay, etc. The detection process and operation process of the above detection methods are relatively troublesome. , unable to adapt to large sample detection

Method used

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  • Aflatoxin enzyme linked immunosorbent assay kit and detecting method
  • Aflatoxin enzyme linked immunosorbent assay kit and detecting method

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preparation example Construction

[0037] In order to better prepare aflatoxin B 1 Antigen, in one embodiment, the aflatoxin B 1 The preparation method of antigen comprises the following steps:

[0038] aflatoxin B 1 Place carboxymethylhydroxylamine hemihydrochloride in pyridine, stir and react at room temperature in the dark for 24 hours to obtain the first reaction solution;

[0039] Freeze-drying the first reaction solution to obtain a solid dry product;

[0040] Dissolve the dried solid with pure water, adjust the pH to 3.0 with hydrochloric acid, extract the precipitate with ethyl acetate, and obtain aflatoxin B after vacuum drying 1 Oxime;

[0041] aflatoxin B 1 Dissolve the oxime in the pyridine solution of KOH, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, and react at a temperature of 70-80°C for 12 hours to obtain the second reaction liquid;

[0042] N-hydroxysuccinimide was added to the second reaction solution, and stirred at room temperature for 2 hours to obtain a third r...

specific Embodiment

[0097] 1. The composition of the kit

[0098]

[0099] Wherein, the aflatoxin B in the present embodiment 1 The coated antigen is prepared by the following preparation method:

[0100] 20 mg of aflatoxin B 1 Place 30 mg of carboxymethylhydroxylamine hemihydrochloride in 2.0 ml of pyridine, stir and react at room temperature in the dark for 24 hours to obtain the first reaction solution;

[0101] Freeze-drying the first reaction solution to obtain a solid dry product;

[0102] Dissolve the dried solid with 10ml of pure water, adjust the pH to 3.0 with 0.2 mol / L hydrochloric acid, extract the precipitate with ethyl acetate, and obtain aflatoxin B after vacuum drying 1 Oxime;

[0103] 10 mg of aflatoxin B 1 Dissolve the oxime in 1ml of 0.1 mol / liter KOH pyridine solution, add 6.5mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), at 70~ reacting at a temperature of 80° C. for 12 hours to obtain a second reaction liquid;

[0104] Add 40 mg of N-hydro...

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Abstract

The invention discloses an aflatoxin enzyme linked immunosorbent assay kit and a detecting method. The aflatoxin enzyme linked immunosorbent assay kit comprises: an enzyme label plate, an aflatoxin B1standard solution, an aflatoxin B1 antibody working solution, an enzyme labeled second antibody working solution, a substrate solution A, a substrate solution B, a stop solution, a concentrated diluent and a concentrated cleaning solution. The substrate solution A is a citric acid-dibasic sodium phosphate buffer solution containing 0.5 mmol / L of hydrogen peroxide urine. The substrate solution B is an ethanol solution of tetramethylbenzidine. The stop solution is 2 mol / L of a sulfuric acid aqueous solution. The enzyme label plate is coated with an aflatoxin B1 coating antigen. The aflatoxin B1coating antigen is obtained by coupling the aflatoxin B1 antigen with bovine serum albumin. Compared with a traditional detecting method, the above detecting method of the kit is relatively simple inboth detecting process and sample processing process, and the detecting process and the operation are both relatively convenient.

Description

technical field [0001] The application relates to the technical field of mycotoxin inspection, in particular to an aflatoxin ELISA kit and detection method. Background technique [0002] Aflatoxin is a toxic metabolite of Aspergillus flavus and Aspergillus parasiticus. Aflatoxin is one of the strongest carcinogens known at present, and its toxicity is greater than that of cyanide and arsenic. At present, there are more than 20 kinds of aflatoxins that have been discovered, and the main aflatoxin that poses a health threat to humans is aflatoxin B. 1 , B 2 , G 1 , G 2 , M 1 and M 2 . Aflatoxins are mainly found in grains, nuts and feed. 1993 Aflatoxin B 1 It is classified as Class I carcinogen by the Cancer Research Institute of the World Health Organization (WHO), which is a class of highly toxic and highly toxic substances. It has been proved to be very harmful to human and animal tissues and organs such as liver and kidney. my country's regulations: aflatoxin B ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543G01N33/569G01N33/577
Inventor 骆鹏杰赵云峰陈霞王紫菲陈银辉李敬光周爽
Owner CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT
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