Aflatoxin M1 immunochromatographic test strip and preparation method thereof

An immunochromatographic test strip and aflatoxin technology, which is applied in analytical materials, measuring devices, instruments, etc., can solve problems such as the advent of aflatoxin M1 immunochromatographic test strips, and achieve the effect of simple operation and high detection sensitivity.

Inactive Publication Date: 2011-06-29
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

Method used

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  • Aflatoxin M1 immunochromatographic test strip and preparation method thereof
  • Aflatoxin M1 immunochromatographic test strip and preparation method thereof
  • Aflatoxin M1 immunochromatographic test strip and preparation method thereof

Examples

Experimental program
Comparison scheme
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Example Embodiment

Example 1: Preparation method of anti-aflatoxin M1 monoclonal antibody

The anti-aflatoxin M1 monoclonal antibody is produced by the hybridoma cell line 2C9 with the deposit number CCTCCNO.C201018, and its preparation method includes the following steps:

1. Obtain hybridoma cell line 2C9 using a two-step screening method

(1) Animal immunity

Six 6-week-old BALB / c mice were purchased and immunized with the commercially available aflatoxin complete antigen AFM1-BSA. After the first immunization, the complete antigen of aflatoxin and the same amount of Freund's complete adjuvant were emulsified, and then injected into multiple points under the skin of the back of the neck of the mouse. The second immunization was carried out 4 weeks later and was emulsified with Freund's incomplete adjuvant and an equivalent amount of aflatoxin, and injected into the abdominal cavity of the mouse. The third immunization and the second immunization are separated by 4 weeks. The immunization method...

Example Embodiment

Example 2-4: Preparation and application of aflatoxin M1 immunochromatographic test strip

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

Example Embodiment

Example 2

The method for preparing aflatoxin M1 immunochromatographic test strip as described above includes the following steps:

(1) Preparation of absorbent pad

Cut the absorbent paper to a length of 16mm and a width of 3mm to obtain an absorbent pad;

(2 Preparation of detection pad

Figure 1 is a front view of the aflatoxin M1 immunochromatographic test strip of the present invention. In the picture: 1 cardboard, 2 absorbent pad; 3 inspection pad; 4 gold label pad; 5 sample pad; 6 quality control line; 7 inspection line.

The aflatoxin M1-bovine serum albumin conjugate AFM1-BSA was prepared into a 0.1mg / mL coating solution A; at a position 15mm away from the upper edge of the nitrocellulose membrane, spray the coating solution A Coated horizontally 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;

The coatin...

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Abstract

The invention belongs to the field of biological detection. An aflatoxin M1 immunochromatographic test strip is characterized by comprising a paper board, wherein a water absorption pad, a detection pad, a gold-labeled pad and a sample pad are attached to one side of the paper board in turn from top to bottom, wherein the adjacent pads are overlapped at joints; the detection pad takes a nitrocellulose membrane as a base pad; the nitrocellulose membrane is provided with a transverse quality control line and a transverse detection line from top to bottom; the detection line is coated with an aflatoxin M1-bovine serum albumin (AFM1-BSA) conjugate; the quality control line is coated with a rabbit anti-mouse polyclonal antibody; a nanogold-labeled aflatoxin M1-resistant monoclonal antibody is transversely sprayed on the gold-labeled pad; and the aflatoxin M1-resistant monoclonal antibody is produced by a hybridoma cell strain 2C9 with the collection number of CCTCCNO.C201018. The test strip is used for detecting aflatoxin M1 and has the characteristics that: the test strip quickly detects the aflatoxin M1, is easy to operate and has high sensitivity.

Description

Aflatoxin M1 immunochromatographic test strip and preparation method thereof technical field 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 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 detecti...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/532
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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