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Chromatographic time-resolved fluorescence kit for simultaneous detection of five mycotoxins and its application

A technology of time-resolved fluorescence and simultaneous detection, which is applied in fluorescence/phosphorescence, analytical materials, measurement devices, etc., and can solve problems such as long detection time, expensive instruments, and complicated sample pretreatment process

Active Publication Date: 2019-08-13
OIL CROPS RES INST 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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  • Chromatographic time-resolved fluorescence kit for simultaneous detection of five mycotoxins and its application

Examples

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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 immunochromatographic time-resolved fluorescent reagent kit for synchronous detection of mixed pollution of five kinds of mycotoxins such as aflatoxin B1 and its application. It includes immunochromatographic time-resolved fluorescent test strips and sample reaction bottles containing lyophilized products of europium-labeled monoclonal antibodies; the fluorescent test strips include a PVC substrate, and one side of the substrate is sequentially pasted with water-absorbing pads from top to bottom , detection pad and sample pad, adjacent pads are overlapped and connected at the junction, the detection pad is based on nitrocellulose membrane, and horizontal quality control lines and 5 detection lines are set from top to bottom, respectively coated with each toxin The bovine serum albumin conjugate and the fumonisin B1 monoclonal antibody are secreted and produced by the hybridoma cell line Fm7A11 with the preservation number CCTCC NO.C201636. It can be used for simultaneous detection of mixed contamination of aflatoxin B1, fumonisin B1, ochratoxin A, zearalenone, and variegated toxins. It is simple, fast and sensitive to operate.

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