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Aflatoxin M1 (AFM1) immunity chromatography test paper strip and preparation method thereof

An immunochromatographic test strip and aflatoxin technology, which can be applied in analytical materials, measuring devices, instruments, etc., can solve the problems such as the advent of aflatoxin M1 immunochromatographic test strips.

Active Publication Date: 2011-11-23
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no immunochromatographic test strip for aflatoxin M1 in the world at present.

Method used

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  • Aflatoxin M1 (AFM1) immunity chromatography test paper strip and preparation method thereof
  • Aflatoxin M1 (AFM1) immunity chromatography test paper strip and preparation method thereof
  • Aflatoxin M1 (AFM1) immunity chromatography test paper strip and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the preparation method of anti-aflatoxin M1 monoclonal antibody

[0052] The anti-aflatoxin M1 monoclonal antibody is produced by the hybridoma cell line 2C9 whose preservation number is CCTCC NO. C201018, and its preparation method comprises the following steps:

[0053] 1. Obtain hybridoma cell line 2C9 by two-step screening method

[0054] (1) Animal immunity

[0055] Six 6-week-old BALB / c mice were purchased and immunized with commercially available aflatoxin complete antigen AFM1-BSA. For the first immunization, the aflatoxin complete antigen was emulsified with an equal amount of Freund's complete adjuvant, and then injected subcutaneously at multiple points on the back of the neck of the mouse. The second immunization was carried out 4 weeks later, and the complete antigen was emulsified with the same amount of aflatoxin in Freund's incomplete adjuvant, and injected intraperitoneally into the mice. The interval between the third immunization ...

Embodiment 2-4

[0069] Example 2-4: Preparation and Application of Aflatoxin M1 Immunochromatographic Test Strip

[0070] The anti-aflatoxin M1 monoclonal antibody used in the following Examples 2-4 is the anti-aflatoxin M1 monoclonal antibody prepared in Example 1.

Embodiment 2

[0072] The preparation method of aflatoxin M1 immunochromatographic test strip comprises the following steps:

[0073] (1) Preparation of absorbent pad

[0074] Cut the absorbent paper to a size of 16mm and 3mm wide to obtain an absorbent pad;

[0075] (2 Preparation of detection pad

[0076] The coating of the detection line:

[0077] Prepare aflatoxin M1-bovine serum albumin conjugate AFM1-BSA into 0.1mg / mL coating solution A; spray coating solution A at a position 15mm away from the upper edge of the nitrocellulose membrane Horizontally coated on a nitrocellulose membrane to obtain a detection line, the coating amount of aflatoxin M1-bovine serum albumin conjugate required per centimeter of the detection line is 75ng, and then dried at 37°C for 15 minutes;

[0078] The coating solution A is: 10 mg of commercially available aflatoxin M1-bovine serum albumin conjugate AFM1-BSA, 1 g of bovine serum albumin, 0.02 g of sodium azide, 0.8 g of sodium chloride, 0.29 g of ten Di...

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Abstract

The invention belongs to the field of biological detection. An AFM1 immunity chromatography test paper strip is characterized in that: the test paper strip comprises a paperboard, an absorbent pad, a detection pad, a colloidal gold pad and a sample pad are sequentially pasted on one side of the paperboard, and adjacent pads overlappingly connect at a joint; the detection pad treats a cellulose nitrate membrane as a base pad, a quality control wire and a detection wire are arranged on the cellulose nitrate membrane in a top-down manner, the detection wire is coated with an AFM1-bovine serum albumin (AFM1-BSA) conjugate, and the quality control wire is coated with a rabbit-anti-mouse polyclonal antibody; and the colloidal gold pad is transversely sprayed with a nanogold labeled AFM1 monoclonal antibody which is generated by a hybridoma cell strain 2C9 with the accession number of CCTCCNO.C201018. The test paper strip which is used for AFM1 detection has the characteristics of rapid detection, simple operation and high sensitivity.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to an aflatoxin M1 immunochromatographic test strip and a preparation method thereof. Background technique [0002] Aflatoxin is mainly a secondary metabolite secreted by Aspergillus flavus and Aspergillus parasiticus, and is a natural toxic compound that can cause various damages to humans and animals. More than 20 types of aflatoxin (AFT) have been discovered, among which aflatoxin B1 is the most toxic and carcinogenic substance. Aflatoxin M1 is a hydroxylated metabolite of aflatoxin B1 and is also a strong carcinogen. When mammals ingest AFB1-contaminated feed, AFM1 is secreted into milk through hydroxylation in vivo. Aflatoxin M1 is present in milk produced by dairy cows fed feed contaminated with aflatoxin B1. Since aflatoxin M1 is quite stable, it cannot be killed by pasteurization, so the detection of aflatoxin M1 should not only detect animal feed raw mater...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/532
Inventor 李培武张道宏张奇张文管笛丁小霞姜俊
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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