Immunity chromatography test strip for synchronously detecting aflatoxin and ochratoxin A mixed pollution, and preparation method and application thereof

An immunochromatographic test paper and aflatoxin technology, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of mixed pollution of food and feed, strong toxicity, possibility of mixed pollution of mycotoxins, etc.

Inactive Publication Date: 2013-09-04
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The two are not only highly toxic, but also have a wide range of pollution. There are many reports on the harm of aflatoxins to food and feed. Ochratoxin A has also been detected in many countries to contaminate food crops in large quantities. In addition, there are data showing that the two are also easily Contamination of the same food crop at the same time, resulting in mixed pollution of food and feed
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 mixed mycotoxin contamination of the same agricultural product is high. Simultaneous and rapid monitoring of mycotoxin mixed contamination in feed

Method used

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  • Immunity chromatography test strip for synchronously detecting aflatoxin and ochratoxin A mixed pollution, and preparation method and application thereof
  • Immunity chromatography test strip for synchronously detecting aflatoxin and ochratoxin A mixed pollution, and preparation method and application thereof
  • Immunity chromatography test strip for synchronously detecting aflatoxin and ochratoxin A mixed pollution, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Obtaining of anti-aflatoxin universal monoclonal antibody and anti-ochratoxin A monoclonal antibody

[0044] 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 with the preservation number CCTCC NO.C201018 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. The specific operation Method: filter mouse ascites with double-layer filter paper, centrifuge at 12000r / min for 15min at 4°C, absorb the supernatant, mix the obtained ascites supernatant with 4 times the volume of acetate buffer, and slowly add n-octanoic acid under stirring, per ml The volume of ...

Embodiment 2

[0065] The preparation method of the immunochromatographic test strip for simultaneous detection of mixed contamination of aflatoxin and ochratoxin A, the steps are as follows:

[0066] (1) Preparation of absorbent pad

[0067] Cut the absorbent paper to a length of 16mm and a width of 4mm to obtain an absorbent pad;

[0068] (2) Preparation of detection pad

[0069] Prepare ochratoxin A-bovine serum albumin conjugate (OTA-BSA) with coating buffer to make a coating solution of 0.4 mg / mL, and place it at a position 15 mm away from the edge of the nitrocellulose membrane, and spray it It is coated on the nitrocellulose membrane to obtain the detection line I, and the coating amount of the required ochratoxin A-bovine serum albumin conjugate (OTA-BSA) on the detection line I per centimeter is 150ng; Toxin 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 the nitrocellulose membrane with a spot spray m...

Embodiment 3

[0092] The preparation method of the immunochromatographic test strip for simultaneous detection of mixed contamination of aflatoxin and ochratoxin A, the steps are as follows:

[0093] (1) Preparation of absorbent pad

[0094] Cut the absorbent paper to a length of 18mm and a width of 3mm to obtain an absorbent pad;

[0095] (2) Preparation of detection pad

[0096] Prepare ochratoxin A-bovine serum albumin conjugate (OTA-BSA) with coating buffer to make a coating solution of 0.4 mg / mL, and spray it at a position 20 mm away from the upper edge of the nitrocellulose membrane. It is coated on nitrocellulose membrane to obtain detection line I, and the coating amount of required ochratoxin A-bovine serum albumin conjugate (OTA-BSA) on every centimeter detection line I is 300ng; Aspergillus flavus Toxin B1-bovine serum albumin conjugate (AFB1-BSA) is prepared into a solution of 0.5 mg / mL with the coating solution, and is coated on the nitrocellulose membrane with a spot spray m...

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Abstract

The invention relates to an immunity chromatography test strip for synchronously detecting aflatoxin and ochratoxin A mixed pollution, and a preparation method and an application thereof. The test strip comprises a cardboard, a water absorption pad, a detection pad, a colloidal gold pad and a sample pad are sequentially pasted on one surface of the cardboard from top to bottom, adjacent pads are connected at the connection in an overlapping manner, the detection pad treats a nitrocellulose membrane as a base pad, the nitrocellulose membrane is provided with a transverse quality control line and detection lines, the detection lines are positioned below the quality control line, the number of the detection lines is two, the detection lines are distributed in an interval manner and are coated with an ochratoxin A-bovine serum albumin conjugate and an aflatoxin B1-bovine serum albumin conjugate respectively, and the quality control line is coated with a rabbit anti-mouse polyclonal antibody; and the colloidal gold pad is transversely sprayed with a nano-gold labeled anti-aflatoxin universal monoclonal antibody and a nano-gold labeled anti-zearalenone monoclonal antibody. The immunity chromatography test strip can be used for synchronously detecting the contents of aflatoxin and ochratoxin A in a sample, and has the characteristics of simple operation, rapidness 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 ochratoxin A, a preparation method and an application thereof. Background technique [0002] Mycotoxins (mycotoxins) are toxic secondary metabolites produced by fungi during their growth. Mycotoxins are usually low-molecular substances, and most mycotoxins are heat-stable substances. Since the discovery of mycotoxins, there are more than 300 kinds of mycotoxins that have been confirmed, among which aflatoxins and ochratoxins are the two most toxic types of toxins. Aflatoxin is listed as a class I carcinogen by the International Agency for Research on Cancer, and ochratoxin A in ochratoxin is listed as a class II carcinogen. The two are not only highly toxic, but also have a wide range of pollution. There are many reports on the harm of aflatoxins to food and feed. O...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
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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