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Immunochromatography time resolution fluorescence kit for synchronously detecting mixed pollution of five types of fungi toxins fungaltoxin of aflatoxin and the like and application

A time-resolved fluorescence and aflatoxin technology, applied in fluorescence/phosphorescence, analytical materials, measurement devices, etc., can solve problems such as high requirements, poor reproducibility, and complicated sample pretreatment process

Active Publication Date: 2017-07-07
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

Thin-layer chromatography is simple and convenient, but has poor reproducibility and low precision; enzyme-linked immunoassay has strong specificity and simple pretreatment, but has a high false positive rate and cannot be used as a confirmation method; liquid chromatography and liquid chromatography-mass spectrometry Good stability and high sensitivity, but the sample pretreatment process is complicated, the detection time is long, the instruments used are expensive, the requirements for the experimental environment and testing personnel are high, and it is difficult to achieve rapid detection

Method used

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  • Immunochromatography time resolution fluorescence kit for synchronously detecting mixed pollution of five types of fungi toxins fungaltoxin of aflatoxin and the like and application

Examples

Experimental program
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Effect test

Embodiment 1

[0040] The acquisition of embodiment 1 aflatoxin B1 monoclonal antibody

[0041] The aflatoxin universal monoclonal antibody is secreted and produced by the hybridoma cell line 3G1 with the deposit number CCTCC NO.C201014. Specifically, it is pre-prepared according to the method reported in the patent with the authorization number ZL201210117614.9. The preparation method is: the obtained hybrid The tumor cell line 3G1 was injected into BALB / c mice pre-treated with Freund's incomplete adjuvant, the ascites of the mice was collected, purified and processed to obtain aflatoxin B1 monoclonal antibody. Among them, the purification method is the octanoic acid-ammonium sulfate method, and the specific operation is: 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, and dilute the supernatant with 4 times the volume Mix acetate buffer solution, slowly add n-octanoic acid while stirrin...

Embodiment 2

[0043] Example 2 Obtaining of Ochratoxin A Monoclonal Antibody

[0044] Ochratoxin A monoclonal antibody is secreted and produced by the hybridoma cell line 1H2 with the deposit number CCTCC NO.C201329. Specifically, it is prepared in advance according to the method reported in the patent application number 201310115921.8. The preparation method is: hybridoma cell line 1H2 Inject into the abdomen of BALB / c mice treated with Freund's incomplete adjuvant in advance, collect the ascites of the mice, and purify to obtain ochratoxin A monoclonal antibody. The purification method is caprylic acid-ammonium sulfate method, and the specific steps are: filter mouse ascites with double-layer filter paper, centrifuge at 12,000 r / min for more than 15 minutes at 4°C, absorb the supernatant, and mix the obtained ascites supernatant with 4 times the volume of vinegar Mix salt buffer solution, add n-octanoic acid slowly under stirring, the volume of n-octanoic acid required per ml of ascites i...

Embodiment 3

[0046] Example 3 Obtaining of Zearalenone Monoclonal Antibody

[0047]The zearalenone monoclonal antibody is secreted and produced by the hybridoma cell line 2D3 with the deposit number CCTCC NO.C201328. Specifically, it is prepared in advance according to the method reported in the patent application number 201310115825.3. The preparation method is as follows: the hybridoma cell line 2D3 was injected into the abdomen of BALB / c mice pre-treated with Freund's incomplete adjuvant, the ascites of the mice was collected, and purified to obtain the zearalenone monoclonal antibody; the purification method is caprylic acid-ammonium sulfate The specific steps are as follows: filter mouse ascites with double-layer filter paper, centrifuge at 12,000 r / min for more than 15 minutes 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, the volume of n-octanoic acid required per milliliter of ascites i...

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Abstract

The invention relates to an immunochromatography time resolution fluorescence kit for synchronously detecting mixed pollution of five types of fungaltoxinfungi toxins of aflatoxin and the like and application. The immunochromatography time resolution fluorescence kit comprises an immunochromatography time resolution fluorescence test paper strip and sample reaction bottles of monoclonal antibody freeze-drying samples containing europium labels, wherein the fluorescence test paper strip comprises a PVC (polyvinyl chloride) substrate; a water absorbing pad, a detection pad and a sample pad are sequentially adhered onto one surface of the substrate; the adjacent pads are overlapped and connected at the connecting part; the detection pad uses a nitrocellulose membrane as a base pad, and is provided with a transverse quality control line and five detection lines from top to bottom to respectively coat the bovine serum albuminaflatoxin B1-BSA conjugate of each toxin; a fumonisin B1 monoclonal antibody is excreted by a hybrid tumor cell strain Fm7A11 with collection number of CCTCC NO.C201636. The immunochromatography time resolution fluorescence kit is applied to synchronously detect the mixed pollution of toxins of aflatoxin B1, fumonisin B1, ochratoxin A, zearalenone and aspergillus versicolor, and has the advantages that the operation is simple and quick, and the sensitivity is high.

Description

technical field [0001] The invention relates to a mycotoxin immunochromatography time-resolved fluorescence kit, in particular to an immunochromatography time-resolved fluorescence kit for synchronously detecting mixed contamination of five mycotoxins such as aflatoxin B1 and its application. Background technique [0002] Mycotoxins are a series of toxic and harmful substances produced by fungi growing and metabolizing in grain, oil or feed. Currently, more than 400 mycotoxins have been found in nature. According to the main toxin-producing bacteria species, mycotoxins can be divided into several categories such as aspergillus toxins (such as aflatoxins, variegated aspergillus toxins, etc.), penicillium toxins and fusarium toxins (such as zearalenone, etc.). Mycotoxins have carcinogenic, teratogenic and mutagenic effects, can cause acute or chronic poisoning of the human body, and seriously endanger human health. Mycotoxins contaminate crops, food and feed, posing a huge th...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/577G01N33/558
CPCG01N21/6408G01N33/558G01N33/577G01N21/8483G01N2021/7786G01N2021/7759G01N33/56961G01N2458/40G01N33/54388G01N2333/38
Inventor 张兆威李培武张奇王督张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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