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Ultra-sensitive aflatoxin B1 enzyme-linked immunosorbent assay kit

A technology for enzyme-linked immunosorbent assay and aflatoxin, which is applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as inability to perform accurate quantification, and achieve the effects of good promotion value, convenient operation and convenient carrying.

Active Publication Date: 2013-04-03
BEIJING PRIMEBIOTEK COMPANY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, aflatoxin B 1 The detection limit of the enzyme-linked immunoassay method can usually only reach 0.1ng / mL, and for lower concentrations of aflatoxin B 1 cannot be accurately quantified

Method used

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  • Ultra-sensitive aflatoxin B1 enzyme-linked immunosorbent assay kit
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  • Ultra-sensitive aflatoxin B1 enzyme-linked immunosorbent assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] A hypersensitive aflatoxin B 1 Preparation of enzyme-linked immunoassay kit

[0076] 1. Preparation of Aflatoxin B 1 standard solution:

[0077] aflatoxin B 1 Prepared from pure product, divided into 6 bottles of 0, 0.1, 0.3, 0.9, 2.7, 8.1ng / mL, sterilized by filtration, divided into packages, and stored at low temperature and protected from light.

[0078] 2. Aflatoxin B 1 Monoclonal Antibody Coated Microplates:

[0079] 1) coated

[0080] Adopt 0.05mol / L pH to be the carbonate buffer of 9.6 and the aflatoxin B of suitable concentration 1 The monoclonal antibody was mixed to make a coating solution, which was loaded on a solid phase carrier, 100 μL per well of a microplate, and incubated at 4°C.

[0081] 2) washing

[0082] Wash with phosphate buffered saline containing 0.05% Tween 20 by volume.

[0083] 3) Protection

[0084] Add the protective solution to the washed solid phase carrier, 150 μL per well, place at 37°C for 1 hour, shake off the protective sol...

example 2

[0100] aflatoxin B 1 Preparation of enzyme-linked immunoassay kit

[0101] 1. Aflatoxin B 1 Fully antigen-coated microplates:

[0102] 1) coated

[0103] Adopt 0.05mol / L pH to be the carbonate buffer of 9.6 and the aflatoxin B of suitable concentration 1 The complete antigen was mixed to make a coating solution, which was loaded on a solid phase carrier, 100 μL per well of a microplate, and incubated at 4°C.

[0104] 2) washing

[0105] Wash with phosphate buffered saline containing 0.05% Tween 20 by volume.

[0106] 3) Protection

[0107] Add the protective solution to the washed solid phase carrier, 150 μL per well, place at 37°C for 1 hour, shake off the protective solution, pat dry on absorbent paper, dry in a 37°C drying oven for 1 hour, and immediately vacuum seal the bag. Put a desiccant in each bag when sealing the bag. Leave the bag for 30 minutes to check for air leaks, and reseal the bag if there is an air leak. After checking that there is no air leakage, ...

Embodiment 3

[0118] Detection of aflatoxin B using the enzyme-linked immunosorbent assay kit prepared in Examples 1-2 of the present invention 1 Methods

[0119] 1. Sample pretreatment:

[0120] Crush the sample into powder, mix it with the sample extract at a ratio of 1:2, sieve and take the supernatant and store it at 4°C, and dilute it with the sample diluent 1:9 to be the solution to be tested when it is used. 20 times.

[0121] 2. Using ultra-sensitive aflatoxin B 1 The enzyme-linked immunoassay kit detects the content of aflatoxin B1 in the sample.

[0122] 1) Take the kit out from 4°C and place it at room temperature for 1 hour. Add 50 μL / well of aflatoxin B1 standard solution or test solution and 50 μL / well of enzyme marker into the coated microwell plate, and mix well for 25 Incubate at ℃ for 30 minutes;

[0123] 2) After washing the above-mentioned microwell plate with washing solution, add 100 μL / well of substrate chromogenic solution, and react in the dark at 25°C for 5 mi...

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Abstract

The invention discloses an ultra-sensitive aflatoxin B1 enzyme-linked immunosorbent assay kit. The kit comprises an aflatoxin B1 standard solution, a solid phase carrier, an enzyme labeling object, a substrate developing solution, a sample dilute solution, a termination solution and a concentration washing solution, wherein the solid phase carrier is a micropore plate; and the enzyme labeling object is an aflatoxin B1 protein coupling object labeled by horseradish peroxide and is coated by an aflatoxin B1 monoclonal antibody, and the substrate developing solution is tetramethyl benzidine (TMB). The invention also discloses a preparation method of the ultra-sensitive aflatoxin B1 enzyme-linked immunosorbent assay kit and a method for detecting aspergillus flavus B1. By using the kit according to the invention to detect aflatoxin B1 the kit has the characteristics of simple operation, high sensitivity, good specificity, good linearity and the like.

Description

technical field [0001] The invention relates to the field of analysis and detection reagents, more specifically, to a method for detecting aflatoxin B 1 The enzyme-linked immunoassay assay kit and its preparation and use methods. Background technique [0002] Aflatoxin (AF) is a highly toxic substance, which has been classified as a Class 1 carcinogen by the Cancer Research Institute of the World Health Organization. Aflatoxins are metabolites of toxin-producing strains of Aspergillus flavus and Aspergillus parasiticus, and are commonly found in moldy grains, feeds and their products. Aflatoxin B 1 , B 2 , G 1 and G 2 It is the main form of aflatoxin in grain and food, of which aflatoxin B 1 most toxic and carcinogenic. AFB 1 is the main one, and its toxicity and carcinogenicity are the greatest, therefore, AFB is usually used in the test 1 as an analysis indicator. [0003] Aflatoxin B 1 The molecular formula is C 17 h 12 o 6 , the chemical structure is as fol...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 时国庆孙清杨敬臣李谷丰
Owner BEIJING PRIMEBIOTEK COMPANY
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