Immunochromatography test strip for synchronously detecting mixed pollution of aflatoxin, ochratoxin A and zearalenone, and preparation method

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

Inactive Publication Date: 2013-07-24
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 immunochromatographic test strips 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 same agricu

Method used

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  • Immunochromatography test strip for synchronously detecting mixed pollution of aflatoxin, ochratoxin A and zearalenone, and preparation method
  • Immunochromatography test strip for synchronously detecting mixed pollution of aflatoxin, ochratoxin A and zearalenone, and preparation method
  • Immunochromatography test strip for synchronously detecting mixed pollution of aflatoxin, ochratoxin A and zearalenone, and preparation method

Examples

Experimental program
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Example Embodiment

[0049] Example 1: Obtainment of anti-aflatoxin universal monoclonal antibody, anti-ochratoxin A monoclonal antibody and anti-zearalenone monoclonal antibody

[0050] a. The anti-aflatoxin universal monoclonal antibody is secreted by the hybridoma cell line 1C11 with the deposit number of CCTCC NO.C201018. Specifically, it is prepared in advance according to the method reported in the patent with the patent application number of CN201010245095.5. The preparation method is: The hybridoma cell line 1C11 was injected into BALB / c mice pretreated with incomplete Freund's adjuvant, the ascites of the mice was collected, and the antibody was purified by the octanoic acid-ammonium sulfate method. Rat ascites was centrifuged at 12000r / min for 15min at 4°C, the supernatant was aspirated, the obtained ascites supernatant was mixed with 4 times the volume of acetate buffer, and n-octanoic acid was slowly added under stirring. The required volume of n-octanoic acid per milliliter of ascites wa...

Example Embodiment

Example 2

The preparation method of the immunochromatographic test strip for simultaneous detection of mixed contamination of aflatoxin, ochratoxin A and zearalenone, the steps are as follows:

(1) Preparation of absorbent pads

Cut the absorbent paper to a length of 16mm and a width of 4mm to get the absorbent pad;

(2) Preparation of detection pad

The coating of the detection line:

The ochratoxin A-bovine serum albumin conjugate (OTA-BSA) was prepared into a coating solution of 0.4 mg / mL with coating buffer, and was sprayed at a position 15 mm away from the upper edge of the nitrocellulose membrane. It is coated on nitrocellulose membrane to obtain detection line I, and the required ochratoxin A-bovine serum albumin conjugate (OTA-BSA coating amount is 160 ng per centimeter of detection line I; Enone-bovine serum albumin conjugate (ZEA-BSA) was prepared into 0.4 mg / mL coating solution with coating buffer, and it was coated on nitrocellulose by spot spraying at a positi...

Example Embodiment

Example 3

[0012] The preparation method of the immunochromatographic test strip for simultaneous detection of mixed contamination of aflatoxin, ochratoxin A and zearalenone as described above comprises the following steps:

[0031] The 0.1 mol / L potassium carbonate aqueous solution is: 13.8 g of potassium carbonate is dissolved in pure water to 1000 mL, and filtered through a 0.22 μm filter membrane; the labeled washing and preservation solution is: 2.0 g polyethylene glycol-20000, 0.2 g of sodium azide, 0.1235 g of boric acid, and purified water to 1000 mL, and filtered through a 0.22 μm membrane.

Cut the absorbent paper to a length of 18mm and a width of 3mm to get the absorbent pad;

(2) Preparation of detection pad

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Abstract

The invention relates to an immunochromatography test strip for synchronously detecting the mixed pollution of aflatoxin, ochratoxin A and zearalenone, and a preparation method. The immunochromatography test strip comprises a paperboard, wherein a water-absorbing pad, a detection pad, a gold label pad and a sample pad are sequentially bonded on one surface of the paperboard from top to bottom, and the adjacent pads are connected at a junction in an overlapping manner; the detection pad takes a nitrocellulose membrane as a base pad, and a transversal quality control line and detection lines are arranged on the detection pad; three detection lines are located below the quality control line, distributed at intervals, and coated with OTA-BSA (ochratoxin A-bovine serum albumin), ZEA-BSA (zearalenone A-bovine serum albumin) and AFB1-BSA (aflatoxin B1-bovine serum albumin) respectively; and the quality control line is coated with a rabbit anti-mouse polyclonal antibody; the gold label pad is transversally spayed with a nanogold labelled anti-aflatoxin universal monoclonal antibody, a nanogold labelled anti-ochratoxin A monoclonal antibody and a nanogold labelled anti-zearalenone monoclonal antibody. The immunochromatography test strip can be used for synchronous detection for aflatoxin, ochratoxin A and zearalenone, as well as is simple and rapid to operate, and high in sensitivity.

Description

technical field [0001] The invention provides an immunochromatographic test strip and a preparation method for synchronously detecting mixed pollution of aflatoxin, ochratoxin A and zearalenone. Background technique [0002] Mycotoxins are metabolites produced by fungi growing in food or feed. If the food and feed are not fully dried at the time of harvest or the temperature or humidity is too high during storage and transportation, it may provide suitable conditions for the growth and reproduction of fungi, thereby causing Mycotoxin contamination. Aflatoxins, ochratoxins and zearalenone are common mycotoxins in grain feed. Aflatoxin is mainly produced by Aspergillus flavus and Aspergillus parasiticus, ochratoxin is mainly produced by various Aspergillus and Penicillium, zearalenone is mainly produced by Fusarium graminearum, three kinds of mycotoxins are mainly produced in grains including wheat, corn, barley , oats, rye, rice and millet, etc., as well as peanuts, beans a...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/544
Inventor 李培武张兆威张奇丁小霞张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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