Quantum dot fluorescent microsphere chromatography kit for simultaneously detecting mixed contamination of aspergillus flavus metabolites cyclopiazonic acid and aflatoxin

A technology of cyclopianilic acid and aflatoxin, which is applied in the field of quantum dot fluorescent microsphere chromatography kits, can solve the problems of large amount of reagents, high requirements on experimental environment, complicated processing procedures, etc., and achieves increased stability and detection. Reproducibility, reduced quenching effect, high sensitivity effect

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

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Problems solved by technology

Thin-layer chromatography is commonly used in the early detection of mycotoxins. This method is easy to operate and can be widely used. However, the amount of reagents used is large, the interference of other components is serious, the accuracy is poor, and it cannot be accurately quantified, and it is harmful to the experimenters and the surrounding environment. Large, not suitable for on-site rapid detection
Precision instrument analysis mainly includes liquid chromatography-mass spectrometry and high-performance liquid chromatography. These methods have high sensitivity and good accuracy, but they are expensive and require a high degree of purification of mycotoxin samples. The sample pretreatment process is cumbersome and expensive. It is difficult to achieve rapid detection due to lack of time and high requirements for the experimental environment

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  • Quantum dot fluorescent microsphere chromatography kit for simultaneously detecting mixed contamination of aspergillus flavus metabolites cyclopiazonic acid and aflatoxin

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1: Obtaining of Anti-Cyclopiazonic Acid Monoclonal Antibody

[0041] 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:

[0042] 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...

Embodiment 2

[0060] Embodiment 2: Obtaining of anti-aflatoxin universal monoclonal antibody

[0061] 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 1C11 was injected intraperitoneally into BALB / c mice treated with incomplete Freund's 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 3

[0064] Example 3: A Quantum Dot Fluorescent Microsphere Chromatography Kit for Simultaneous Detection of Cyclopiazonic Acid and Aflatoxin Mixed Pollution and Its Application

[0065] Simultaneous detection of cyclopiazonic acid and aflatoxin mixed contamination Quantum dot fluorescent microsphere chromatography kit, including chromatography test strips, anti-cyclopianic acid monoclonal antibody lyophilized product labeled with quantum dot fluorescent microspheres , Sample reaction bottle of freeze-dried product of anti-aflatoxin universal monoclonal antibody labeled with quantum dot fluorescent microspheres, sample diluent and sample diluent pipette. The chromatographic test strips are as figure 1 As shown, including cardboard, one side of the cardboard is pasted with water-absorbing pad 1, testing pad 2 and sample pad 3 sequentially, and adjacent pads are overlapped and connected at the joints, and the overlapping length is 1 mm. The nitrocellulose membrane is used as the ba...

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Abstract

The invention relates to a quantum dot fluorescent microsphere chromatography kit for simultaneously detecting mixed contamination of aspergillus flavus metabolites cyclopiazonic acid and aflatoxin. The kit comprises a fluorescent chromatography test strip, and a sample reaction bottle containing an anti-cyclopiazonic acid monoclonal lyophilized antibody labeled by the quantum dot fluorescent microsphere and an anti-aflatoxin monoclonal lyophilized antibody labeled by the quantum dot fluorescent microsphere. The anti-cyclopiazonic acid monoclonal antibody is secreted by a hybridoma cell strainYTT-2 with a preservation number of CCTCC NO. C201871. The kit of the present invention can be used for simultaneously detecting cyclopiazonic acid and aflatoxin and has simple and rapid operation and high sensitivity.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a quantum dot fluorescent microsphere chromatography kit for synchronously detecting mixed pollution of cyclopiazonic acid and aflatoxin and an application thereof. Background technique [0002] Mycotoxins are a type of highly toxic substances produced by harmful fungi. They have been found to widely contaminate my country's main grains and oils, mainly including rice, peanuts, corn, and wheat. Mycotoxins can contaminate grain and oil planting, harvesting, transportation, and storage processes, and enter the food chain to endanger life, health and safety. They are listed by the World Health Organization and the Food and Agriculture Organization of the United Nations as the top three causes of foodborne diseases. In order to ensure the quality and safety of grain and oil and promote the high-quality development of the grain and oil industry, my country has formulate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N33/58G01N33/533
CPCG01N33/558G01N33/577G01N33/582G01N33/585G01N33/588G01N33/533
Inventor 张兆威李培武张奇李慧张文胡小风
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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