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Immunochromatographic time resolved fluorescence kit for synchronously detecting aflatoxins, and application of kit

A time-resolved fluorescence and aflatoxin technology, used in biological testing, fluorescence/phosphorescence, analytical materials, etc., can solve problems such as a variety of mycotoxins contamination, and achieve high sensitivity and specificity.

Active Publication Date: 2019-08-09
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

At the same time, the same agricultural product is likely to be contaminated by multiple mycotoxins, so there is an urgent need for a detection technology that can simultaneously detect multiple mycotoxins to achieve simultaneous and rapid monitoring of mixed mycotoxin contamination in food and feed

Method used

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  • Immunochromatographic time resolved fluorescence kit for synchronously detecting aflatoxins, and application of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1: Obtaining of anti-aflatoxin universal monoclonal antibody

[0044] The universal anti-aflatoxin monoclonal antibody is produced by the secretion of the hybridoma cell line 1C11 with the preservation number CCTCC NO.C201013, specifically according to the method reported in the patent application number 201010245095.5. The preparation method is: the obtained hybridoma cells Strain YTT-2 was intraperitoneally injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, and the ascites of the mice was collected, purified and treated to obtain a universal anti-aflatoxin monoclonal antibody. Wherein, the purification method is octanoic acid-ammonium sulfate method, and the specific operation is as follows: the ascites is taken out from the -20°C refrigerator and thawed at room temperature. Filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C, 12000r / min for more than 15min, absorb the supernatant, mix...

Embodiment 2

[0046] Embodiment 2: Obtaining of anti-aspergillus versicolor monoclonal antibody

[0047] The anti-aspergillus versicolor monoclonal antibody is secreted and produced by the hybridoma cell line 1C11 with the deposit number CCTCC NO.C2013187, specifically prepared in advance according to the method reported in the patent application number 201410115952.8. The preparation method is: the obtained hybridoma cells Strain ST03 was intraperitoneally injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, and the ascites of the mice was collected, purified and treated to obtain anti-Varistoxin monoclonal antibodies. Wherein, the purification method is octanoic acid-ammonium sulfate method, and the specific operation is as follows: the ascites is taken out from the -20°C refrigerator and thawed at room temperature. Filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C, 12000r / min for more than 15min, absorb the supernat...

Embodiment 3

[0049] Example 3: Obtaining of Anti-Cyclopiazonic Acid Monoclonal Antibody

[0050] The anti-cyclopianic acid monoclonal antibody is produced by the hybridoma cell line YTT-2 with the preservation number CCTCC NO.C C201871. details as follows:

[0051] Cyclopiazonic acid monoclonal antibody hybridoma cell line YTT-2 was intraperitoneally injected into BALB / c mice treated with incomplete Freund's adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method , the specific operation is: filter mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C, 12000r / min for more than 15min, absorb the supernatant, mix the supernatant with 4 times the volume of acetate buffer, and stir While slowly adding n-octanoic acid, the volume of n-octanoic acid required per milliliter of ascites is 30-35 μL, mix at room temperature for 30-60 min, let stand at 4°C for more than 2 h, then centrifuge at 1...

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Abstract

The invention discloses an immunochromatographic time resolved fluorescence kit for synchronously detecting aflatoxins, and an application of the kit. The kit comprises an immunochromatographic time resolved fluorescence test strip and a sample reaction bottle containing europium-labeled monoclonal antibody freeze-dried samples, wherein the immunochromatographic time resolved fluorescence test strip comprises a bottom plate; a water absorption pad, a detection pad and a sample pad are adhered to the adhesive surface of the bottom plate in sequence from top to bottom; the adjacent pads are connected in an overlapped mode at the joint; the detection pad takes a nitrocellulose membrane as a base pad; a transverse quality control line and a detection line are arranged on the nitrocellulose membrane from top to bottom; the quality control line is coated with a rabbit anti-mouse polyclonal antibody; the detection line is coated with protein conjugates of the toxins; and an anti-cyclopiazonicacid monoclonal antibody is secreted by a hybridoma cell strain YTT-2 with a collection number of CCTCC NO.C201871. The kit can be used for synchronously detecting the content of the aflatoxins, sterigmatocystins and cyclopiazonic acids in the samples, and has the characteristics of simplicity in operation, quickness and high sensitivity.

Description

[0001] Technical field: [0002] The invention belongs to the field of biological detection, and in particular relates to an immunochromatographic time-resolved fluorescent kit for synchronous detection of Aspergillus flavus mycotoxins and an application thereof. Background technique [0003] Aspergillus flavus is a common saprophytic fungus with a wide growth temperature range, easy to grow and reproduce in an environment with high temperature and humidity, and can survive and grow and reproduce under extreme environmental conditions. After Aspergillus flavus infects agricultural products such as peanuts, corn, and tree nuts, it can produce a variety of toxic mycotoxins, causing health problems for humans and animals. [0004] Aflatoxins are a group of structurally similar secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus. They are a group of compounds with difuran ring and coumarin as the basic structure. They are widely found in grains, feed a...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/577G01N33/58G01N33/533G01N21/64
CPCG01N33/558G01N33/577G01N33/582G01N33/533G01N21/6408G01N21/6428
Inventor 张奇李培武王督汪雪芳张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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