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Immunochromatographic test strip for synchronously detecting mixed pollution of aflatoxin and zearalenone, preparation method and application thereof

A technology of zearalenone and immunochromatographic test paper, which is applied to measuring devices, analytical materials, instruments, etc., can solve the problems of high incidence of mycotoxins and contamination of various mycotoxins

Inactive Publication Date: 2013-08-21
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, most of the existing colloidal gold immunochromatographic test strips used for mycotoxin detection can only detect one mycotoxin
However, due to the scattered planting methods of small farmers in my country, the incidence of mycotoxins in agricultural products is high, and the possibility of the same agricultural product being contaminated by multiple mycotoxins is high. Simultaneous and rapid monitoring of mycotoxin mixed contamination in and feed

Method used

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  • Immunochromatographic test strip for synchronously detecting mixed pollution of aflatoxin and zearalenone, preparation method and application thereof
  • Immunochromatographic test strip for synchronously detecting mixed pollution of aflatoxin and zearalenone, preparation method and application thereof
  • Immunochromatographic test strip for synchronously detecting mixed pollution of aflatoxin and zearalenone, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1: Obtaining of anti-aflatoxin universal monoclonal antibody and anti-zearalenone monoclonal antibody

[0046] a. The universal anti-aflatoxin monoclonal antibody is produced by the secretion of the hybridoma cell line 1C11 with the preservation number CCTCC NO.C201018, specifically according to the method reported in the patent application number CN201010245095.5. The preparation method is as follows: The hybridoma cell line 1C11 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. Rat ascites, 4°C, centrifuged at 12000r / min for 15min, sucked the supernatant, mixed the obtained ascites supernatant with 4 times the volume of acetate buffer, slowly added n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites was 33μL, mix at room temperature for 30min, stand at 4°C for 2h, then cent...

Embodiment 2

[0066] The preparation method of the immunochromatographic test strip for synchronously detecting two mycotoxins of aflatoxin and zearalenone, the steps are as follows:

[0067] (1) Preparation of absorbent pad

[0068] Cut the absorbent paper to a length of 16mm and a width of 4mm to obtain an absorbent pad;

[0069] (2) Preparation of detection pad

[0070] The coating of the detection line:

[0071] Prepare zearalenone-bovine serum albumin conjugate (ZEA-BSA) with coating buffer to prepare a coating solution of 0.4 mg / mL, and spray it at a position 15 mm away from the edge of the nitrocellulose membrane. It is coated on the nitrocellulose membrane to obtain the detection line I, and the coating amount of required zearalenone-bovine serum albumin conjugate (ZEA-BSA) on the detection line I per centimeter is 200ng; Aflatoxin B1-bovine serum albumin conjugate (AFB1-BSA) is prepared into a solution of 0.25 mg / mL with coating buffer, and it is coated on nitrocellulose with a ...

Embodiment 3

[0094] The preparation method of the immunochromatographic test strip for synchronously detecting two mycotoxins of aflatoxin and zearalenone, the steps are as follows:

[0095] (1) Preparation of absorbent pad

[0096] Cut the absorbent paper to a length of 18mm and a width of 3mm to obtain an absorbent pad;

[0097] (2) Preparation of detection pad

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Abstract

The invention relates to an immunochromatographic test strip for synchronously detecting mixed pollution of aflatoxin and zearalenone, a preparation method and application thereof. The test strip includes a paperboard. An absorbent pad, a detection pad, a gold labeled pad and a sample pad are sticked on one side of the paperboard from top to bottom. Adjacent pads are in overlapping connection at joints. The detection pad adopts a nitrocellulose membrane as a base pad, which is provided with a transverse control line and detection lines. The two detection lines in internal distribution are positioned below the quality control line, and are respectively coated with a zearalenone-bovine serum albumin conjugate and an aflatoxin B1-bovine serum albumin conjugate. The quality control line is coated with a rabbit antimouse polyclonal antibody. The gold labeled pad is horizontally sprayed with a nanogold labeled anti-aflatoxin universal monoclonal antibody and a nanogold labeled anti-zearalenone monoclonal antibody. The immunochromatographic test strip can be used for simultaneously detecting the content of fungaltoxins aflatoxin and zearalenone in a sample, and has the characteristics of simple operation, rapidity, and high sensitivity.

Description

technical field [0001] The invention relates to a mycotoxin immunochromatographic test strip, in particular to an immunochromatographic test strip for synchronously detecting mixed contamination of aflatoxin and zearalenone, a preparation method and an application thereof. Background technique [0002] Aflatoxins and zearalenone are harmful substances present in grains and are secondary metabolites produced by fungi that contaminate grains. Grain and feed are not sufficiently dried during harvesting or the temperature or humidity is too high during storage and transportation, which may provide suitable conditions for the growth and reproduction of fungi, resulting in mycotoxin contamination. Aflatoxins can strongly damage the liver tissue of humans and animals, and in severe cases can lead to liver cancer or even death; zearalenone has estrogen-like effects, can cause acute and chronic poisoning in animals, cause abnormal reproductive function and even death in animals. Tox...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/531
Inventor 李培武李鑫张奇丁小霞张文张兆威
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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