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Magnetic immuno-chromatographic kit for detecting aflatoxin M1 (AFM1) and preparation method

A magnetic immunochromatography and aflatoxin technology, which is applied in the direction of testing food, measuring devices, analyzing materials, etc., can solve the problems of low sensitivity and low result accuracy, and achieve the effect of high detection sensitivity and simple operation.

Inactive Publication Date: 2015-04-08
BEIJING BIOMASION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since the result is usually observed by human eyes, it is easily affected by the subjective judgment of the observer, the sensitivity is low, and the accuracy of the result is not high.

Method used

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  • Magnetic immuno-chromatographic kit for detecting aflatoxin M1 (AFM1) and preparation method
  • Magnetic immuno-chromatographic kit for detecting aflatoxin M1 (AFM1) and preparation method
  • Magnetic immuno-chromatographic kit for detecting aflatoxin M1 (AFM1) and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: preparation kit of the present invention

[0036] I. Preparation of test strips:

[0037] (1) Treatment of reaction pad: Dilute AFM1-BSA (Sigma-Aldrich, USA, product number A6412) with coating buffer (0.01M PBS solution containing 2% BSA, pH 7.2, pass through a 0.22 μm filter membrane) 0.5 mg / mL, goat anti-mouse IgG antibody (Shanghai Jieyi Biotechnology Co., Ltd., product number JY-QT03) was diluted to 1 mg / mL, and the two Spray it on the nitrocellulose membrane at an interval of 0.8 cm, dry it at 37°C, add a desiccant and store it for later use.

[0038] (2) Treatment of sample pad 2: The sample pad treatment liquid is 0.01M PBS buffer solution with pH 7.2 containing 3% casein and 0.1% Tween-20. Put a 1cm wide sample pad into an appropriate amount of sample pad treatment solution, soak it for 1 hour, take it out, and dry it at 30°C for 8 hours for later use.

[0039] (3) Assembly and cutting of magnetic immunochromatography test strips

[0040] Assem...

Embodiment 2

[0047] Embodiment 2 kit standard curve of the present invention is established and the recovery rate is measured

[0048] 1. Establishment of kit standard curve

[0049] Take an appropriate amount of AFM1 standard substance (Sigma-Aldrich, USA, product number 46319-U) and dilute it to 3.33, 1.11, 0.37, 0.12, 0.04 and 0.01 ng / mL with AFM1-free milk (provided by Beijing Physical and Chemical Analysis and Testing Center), Mix with an equal volume of PBST solution (0.01M pH7.2 PBS containing 1% Triton X-100) respectively, take 100 μL of the above mixed solution and 2 μL superparamagnetic composite particle-labeled AFM1 antibody solution and add it to the test solution in Example 1 In the paper strip, after waiting for 20 minutes to react, insert the test strip into the card slot of the magnetic reader (Quantum Design Company, USA, product number 8094-600-01), and use the magnetic reader to read the test line and the quality control line. MAR value, the results are shown in Table ...

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Abstract

The invention relates to a magnetic immuno-chromatographic kit for detecting aflatoxin M1 (AFM1). The magnetic immuno-chromatographic kit for detecting AFM1 comprises at least one test strip and the solution of a superparamagnetic composite particle labeled AFM1 antibody, wherein the test strip is assembled by adhering a reaction pad, a sample pad 1, a sample pad 2 and an absorbent pad to a base board in sequence in a mutual staggered way and covering the pads with a transparent plastic film, the reaction pad is pre-coated with an AFM1 protein conjugate detection line and an AFM1 antibody combined anti-antibody quantity control line, and the superparamagnetic composite particle labeled AFM1 antibody is a polymer which is formed by combining AFM1 antibody with superparamagnetic composite particles by peptide bonds covalently. The invention further relates to a preparation method of the magnetic immuno-chromatographic kit and application of the magnetic immuno-chromatographic kit to detection of AFM1 in dairy products. The kit is used for immuno-chromatographic detection by taking magnetic particles as labels and has the advantages of high detection speed, convenience in operation, wide linear range and high sensitivity.

Description

technical field [0001] The invention belongs to the field of food detection. Specifically, the present invention relates to a magnetic immunochromatographic kit for detecting aflatoxin M1, its preparation method and the use of the kit for detecting aflatoxin M1. Background technique [0002] Aflatoxins are mainly a group of highly toxic and carcinogenic secondary metabolites produced by Aspergillus flavs, Aspergillus parasiticus and Aspergillus nonius. Aflatoxin M1 (Aflatoxin M1, AFM1) is a food or feed contaminated by aflatoxin B1 (Aflatoxin B1, AFB1), which is catalyzed by hepatic microsomal monooxygenase in the body, through cytochrome P450. Regulatory effect, the terminal furan ring C-10 is hydroxylated to generate. Since it mainly exists in dairy products, the daily consumer products of human beings, especially infants who are more vulnerable to toxin damage, the pollution of AFM1 has aroused widespread concern in the society. In 2002, the level of carcinogenicity of...

Claims

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Application Information

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IPC IPC(8): G01N33/02G01N33/577
Inventor 武会娟管笛亢子佳贾迪
Owner BEIJING BIOMASION TECH
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