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

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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Example Embodiment

[0040] Example 1 Obtaining aflatoxin B1 monoclonal antibody

[0041] The universal monoclonal antibody to aflatoxin is secreted and produced by the hybridoma cell line 3G1 with the deposit number CCTCC NO.C201014, and is specifically prepared in advance according to the method reported in the patent with the grant number ZL201210117614.9. The preparation method is: The tumor cell line 3G1 was injected with BALB / c mice pre-treated with Freund’s incomplete adjuvant, and the ascites of the mice were collected and purified to obtain aflatoxin B1 monoclonal antibody. Among them, the purification method is the caprylic acid-ammonium sulfate method, the specific operation is: filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4℃, 12000r / min for more than 15 minutes, aspirate the supernatant, and mix the supernatant with 4 times the volume Acetate buffer solution mixed, slowly add n-octanoic acid while stirring, the volume of n-octanoic acid requi...

Example Embodiment

[0043] Example 2 Obtaining of Monoclonal Antibodies to Ochratoxin A

[0044] The ochratoxin A monoclonal antibody is secreted and produced by the hybridoma cell line 1H2 with the deposit number CCTCC NO.C201329, and is specifically prepared in advance according to the method reported in the patent application number 201310115921.8. The preparation method is: the hybridoma cell line 1H2 It was injected into the abdomen of BALB / c mice pretreated with Freund's incomplete adjuvant, and the ascites of the mice were collected and purified to obtain ochratoxin A monoclonal antibody. The described purification method is the caprylic acid-ammonium sulfate method, and the specific steps are: filter the mouse ascites with double-layer filter paper, centrifuge at 12000r / min for more than 15 minutes at 4°C, absorb the supernatant, and mix the resulting ascites serum with 4 times the volume of vinegar Mix acid salt buffer, slowly add n-octanoic acid under stirring, the volume of n-octanoic aci...

Example Embodiment

[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, and is specifically prepared in advance according to the method reported in the patent application number 201310115825.3. The preparation method is: 2D3 was injected into the abdomen of BALB / c mice pretreated with Freund’s incomplete adjuvant, and the ascites of the mice were collected and purified to obtain the zearalenone monoclonal antibody; the purification method is caprylic acid-ammonium sulfate Method, the specific steps are: filter the mouse ascites with double-layer filter paper, centrifuge at 12000r / min for more than 15 minutes at 4℃, aspirate the supernatant, mix the resulting ascites serum with 4 times the volume of acetate buffer, and slowly add under stirring Octanoic acid, the required volume of n-octanoic acid per milliliter of ascites is 30-35μL, mix at room tem...

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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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