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Time-resolved fluorescence kit for detecting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously

A time-resolved fluorescence and fusarium enol technology, applied in measuring devices, analytical materials, instruments, etc., can solve the problems of complex pretreatment steps and high cost of sample detection, and achieve the effect of fast detection, high sensitivity and simple operation

Active Publication Date: 2020-02-18
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

These instrument analysis methods have good stability, high sensitivity and good accuracy, but the pretreatment steps are complicated and the cost of sample detection is high

Method used

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  • Time-resolved fluorescence kit for detecting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously
  • Time-resolved fluorescence kit for detecting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously
  • Time-resolved fluorescence kit for detecting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1 Obtaining of anti-diacetylbristol monoclonal antibody

[0045] The anti-diacetyriferinol monoclonal antibody is produced by the secretion of the hybridoma cell line DAS5G11E7 with the preservation number CCTCC NO.C201881, and the preparation method is as follows:

[0046] The hybridoma cell line DAS5G11E7 was 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. Rat ascites, 4°C, centrifuge at 12000r / min for more than 15min, absorb the supernatant, mix the obtained ascites supernatant with 4 times the volume of acetate buffer, slowly add n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites 30-35μL, mix at room temperature for 30-60min, and stand at 4℃ for more than 2h. Centrifuge at 12000r / min at 4°C for more than 30min, discard the precipitate, filter the obtained supernatant with...

Embodiment 2

[0067] Embodiment 2 Preparation of anti-aflatoxin B1 monoclonal antibody

[0068] The anti-aflatoxin B1 monoclonal antibody is secreted and produced by the hybridoma cell line 1C11 with deposit number CCTCC NO.C201013, specifically prepared in advance according to the method reported in the patent application number 201010245095.5. The specific preparation method is as follows: pre-treat BALB / c mice with Freund's incomplete adjuvant, inject the hybridoma cell line 1C11 into the abdomen of the mice, collect ascites after about a week, and purify by caprylic acid-ammonium sulfate method to obtain anti Aflatoxin B1 monoclonal antibody. Specific purification steps: Filter the mouse ascites through double-layer filter paper, centrifuge at 12000r / min, 4°C for 15min to collect the supernatant, aspirate the supernatant into four times the volume of acetate buffer, add n-octanoic acid slowly while stirring, Until the final volume of n-octanoic acid is 30-35 μL / mL, stir and mix at room...

Embodiment 3

[0070] The preparation of embodiment 3 anti-varietatin monoclonal antibodies

[0071] The anti-varietatin monoclonal antibody is secreted and produced by hybridoma cell line ST03 with deposit number CCTCC NO.C2013187, specifically prepared in advance according to the method reported in the patent application number 201410115952.8. The specific preparation method is as follows: pre-treat BALB / c mice with Freund's incomplete adjuvant, inject the hybridoma cell line ST03 into the abdomen of the mice, collect the ascites after about a week, and purify it by the octanoic acid-ammonium sulfate method to obtain the anti Versicolor monoclonal antibody. Specific purification steps: Filter the mouse ascites through double-layer filter paper, centrifuge at 12000r / min, 4°C for 15min to collect the supernatant, aspirate the supernatant into four times the volume of acetate buffer, add n-octanoic acid slowly while stirring, Until the final volume of n-octanoic acid is 30-35 μL / mL, stir and...

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Abstract

The invention discloses a time-resolved fluorescence kit for detecting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously. The time-resolved fluorescence kit comprises an immunochromatographic time-resolved fluorescent test strip and a sample reaction bottle containing an europium-marked monoclonal antibody resisting diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin, whereina water absorption pad, a test pad and a sample pad are arranged at one side of the immunochromatographic time-resolved fluorescent test strip in sequence from top to bottom, adjacent pads overlap andare connected at the joint, the detection pad uses a nitrocellulose membrane as a base pad, a quality control line and three detection lines are transversely arranged on the nitrocellulose membrane from top to bottom, the quality control line wraps a rabbit anti-mouse polyclonal antibody, and the three detection lines respectively wrap toxin-protein conjugates respectively. The kit can rapidly detect the three fungal toxins like diacetoxyscirpenol, aflatoxin B1 and sterigmatocystin simultaneously.

Description

technical field [0001] The invention provides a mycotoxin time-resolved fluorescence immunochromatography kit, specifically a time-resolved method for synchronous detection of mixed contamination of sarcoenol diacetate (snake toxin), aflatoxin B1, and aspergillus versicolor Fluorescence kit and preparation method. Background technique [0002] Diacetoxyscirpenol (4,15-diacetoxyscirpenol, DAS) is a trichothecene toxin produced by Fusarium spp. DAS mainly pollutes grains and feed. It has toxic effects on animal bone marrow, brain, heart, lymph, testis, thymus, nerve cells, etc. It can also cause gastroenteritis, antibody reduction, eye and body cavity edema and other damage. Aflatoxins are a class of toxic secondary metabolites produced by Asperillus flavus and Asperg illusparasiticus infecting crops such as grain and oil, mainly including six types: B1, B2, G1, G2, M1, and M2. The toxicity of aflatoxin B1 is 10 times that of potassium cyanide and 68 times that of arsenic. I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/533
CPCG01N33/577G01N33/558G01N33/533G01N33/54388G01N33/56961G01N33/582
Inventor 李培武李慧姜俊张文张奇
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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