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Immunochromatography time resolution fluorescence kit for synchronization detection of mixed pollution of fumonisin B1 and other four fungaltoxin and application thereof

A time-resolved fluorescence and fumonisin technology, applied in fluorescence/phosphorescence, measuring devices, analytical materials, etc., can solve problems such as low sensitivity, high incidence of mycotoxins, and inability to perform quantitative detection, achieving good specificity and sensitivity high effect

Active Publication Date: 2017-07-11
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

However, due to the low sensitivity and the inability to carry out quantitative detection, it cannot meet the needs of rapid quantitative detection in on-site rapid detection.
However, the scattered planting methods of small farmers in my country lead to a high incidence of mycotoxins in agricultural products, and the risk of mixed contamination is greater. Therefore, there is an urgent need for detection technologies that can quickly and simultaneously detect mixed contamination of various mycotoxins to achieve food and food. Simultaneous and Rapid Monitoring of Mixed Contamination of Mycotoxins in Feed

Method used

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  • Immunochromatography time resolution fluorescence kit for synchronization detection of mixed pollution of fumonisin B1 and other four fungaltoxin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Acquisition of aflatoxin B1 monoclonal antibody

[0040] The aflatoxin B1 monoclonal antibody 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 authorization number ZL201210117614.9. The preparation method is: the obtained hybridoma The tumor cell line 3G1 was injected into BALB / c mice pretreated with incomplete Freund's adjuvant, and the ascites of the mice was collected and purified to obtain aflatoxin B1 monoclonal antibody. Among them, the purification method is octanoic acid-ammonium sulfate method, and the specific operation is as follows: filter the mouse ascites with double-layer filter paper, centrifuge the filtered ascites at 4°C for more than 15 minutes at 12000 r / min, absorb the supernatant, mix the supernatant with 4 times the volume Acetate buffer solution was mixed, and n-octanoic acid was slowly added w...

Embodiment 2

[0042] Example 2 Acquisition of Ochratoxin A Monoclonal Antibody;

[0043] The ochratoxin A monoclonal antibody is secreted and produced by the hybridoma cell line 1H2 with the deposit number of CCTCC NO.C201329, and is specifically prepared in advance according to the method reported in the patent with the application number of 201310115921.8. The preparation method is: the hybridoma cell line 1H2 It was injected into the abdomen of BALB / c mice previously treated with incomplete Freund's adjuvant, and the ascites of the mice was collected and purified to obtain ochratoxin A monoclonal antibody. The purification method is octanoic acid-ammonium sulfate method, and the specific steps are: filtering mouse ascites fluid with double-layer filter paper, centrifuging at 12000 r / min for more than 15 min at 4°C, sucking the supernatant, and mixing the obtained ascites supernatant with 4 times the volume of vinegar. Mix with salt buffer, slowly add n-octanoic acid with stirring, the vo...

Embodiment 3

[0045] The acquisition of embodiment 3 zearalenone monoclonal antibody

[0046] 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 with the application number 201310115825.3. The preparation method is: the hybridoma cell line 2D3 was injected into the abdomen of a BALB / c mouse pretreated with incomplete Freund's adjuvant, and the ascites of the mouse was collected and purified to obtain a zearalenone monoclonal antibody;

[0047] The purification method is octanoic acid-ammonium sulfate method, and the specific steps are: filtering mouse ascites fluid with double-layer filter paper, centrifuging at 12000 r / min for more than 15 min at 4°C, sucking the supernatant, and mixing the obtained ascites supernatant with 4 times the volume of vinegar. Mix with salt buffer, slowly add n-octanoic acid with stirring, the v...

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Abstract

The present invention relates to an immunochromatography time resolution fluorescence kit for synchronization detection of mixed pollution of fumonisin B1 and other four fungaltoxin and application thereof. The immunochromatography time resolution fluorescence kit comprises an immunochromatography time resolution fluorescence test paper strip and a sample reaction bottle containing europium-labeled freeze-dried products of all monoclonal antibodies; the immunochromatography time resolution fluorescence test paper strip includes a PVC substrate, a water absorbent pad, a testing pad and a sample pad which are respectively pasted on one side of the PVC substrate from top to bottom, the testing pad is provided with a nitrocellulose membrane as a base pad, the nitrocellulose membrane is provided with a horizontal quality control line and four detection lines from top to bottom, the four detection lines are respectively coated with bovine serum albumin conjugates of all the fungaltoxin, and a fumonisin B1 monoclonal antibody is secreted by hybridoma cell line Fm7A11 with preservation number of CCTCCNO.C201636. The immunochromatography time resolution fluorescence kit can be used for synchronization detection of content of aflatoxin B1, ochratoxin A, zearalenone and the fumonisin B1, and has the characteristics of simple operation, rapidness and high sensitivity.

Description

technical field [0001] The invention relates to a time-resolved fluorescent kit for mycotoxin immunochromatography, in particular to an immunochromatographic time-resolved fluorescent reagent for simultaneous detection of mixed contamination of aflatoxin B1, ochratoxin A, zearalenone and fumonisin B1 Box, method of preparation and use thereof. Background technique [0002] Mycotoxins are a class of highly toxic chemicals, which are toxic metabolites produced by filamentous fungi under suitable environmental conditions. Mycotoxins widely contaminate corn, wheat, rice, peanuts and other cereal crops and oil crops, feed and other agricultural products. Aflatoxin B1 is a class of toxic secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus. It is highly toxic and highly carcinogenic, teratogenic and mutagenic, and is extremely harmful to humans and animals. Ochratoxin A is mainly produced in some toxin-producing strains of Aspergillus and Penicillium,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/64
CPCG01N21/6486G01N33/577
Inventor 李培武张兆威王督唐晓倩张奇张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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