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Time-resolved fluorescence kit for the simultaneous detection of fusarium diacetate, aflatoxin b1, and versicolor

A time-resolved fluorescence and fusarium enol technology, which is applied in measuring devices, analytical materials, instruments, etc., can solve the problems of complex pre-processing steps and high cost of sample detection, and achieve the effect of high sensitivity, good specificity and simultaneous detection

Active Publication Date: 2021-09-21
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 the simultaneous detection of fusarium diacetate, aflatoxin b1, and versicolor
  • Time-resolved fluorescence kit for the simultaneous detection of fusarium diacetate, aflatoxin b1, and versicolor
  • Time-resolved fluorescence kit for the simultaneous detection of fusarium diacetate, aflatoxin b1, and versicolor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Anti-Acetyllacin Sickle Hylene Alcohol monoclonal antibody

[0045] Anti-acetyl straw sumftone alkanol monoclonal antibody is produced by the secretion of hybridoma cell line DAS5G11E7, which is collected as CCTCN No. C201881, and the preparation method is:

[0046] The hybridoma cell line DAS5G11E7 was injected in advance using a Balb / C mouse, which was used in advance, collecting the ascites of the mouse, purified the antibody using a octanoic-ammonium sulfate method, and the specific operation is: filtering with a double filter paper. The mouse abdomen, 4 ° C, 12000 r / min centrifuged for 15 min, absorb the supernatant, mixed with 4x volume of acetate buffer in the abdomen water, slowly add orthodacosilic acid, and the ytothic acid volume required for per millistened ascites It was 30-35 μL, mixed at room temperature for 30-60 min, and left 4 ° C for 2 hours. 12000 r / min, centrifuged at 4 ° C for 30 min, discarded, and after filtration of the obtained supe...

Embodiment 2

[0067] Example 2 Preparation of monoclonal antibodies against yellow bells B1

[0068] The anti-xantholicin B1 monoclonal antibody is subjected to hybridoma cell line 1C11, which is prepared as CCTCC NO. C201013, and is pre-prepared in advance according to the method reported in the patents of Patent Application No. 201010245095.5. The specific preparation method is as follows: BALB / C mice were treated in advance, and the hybridoma cell line 1C11 was injected into the abdomen of the mouse. After about one week, ascites were collected, purified by hydrazine-ammonium sulfate was purified. Ammyxin B1 monoclonal antibody. Specific purification steps: mouse deposit filtrate filter paper, 12000 r / min, 4 ° C, 15 min centrifugal collected supernatant, absorbed into a quadroyroid of acetate buffer, slowly add orthodaculipine while stirring while stirring. To the endic acid end volume of 30 to 35 μl / ml, stirred at room temperature for 30 to 60 min, and then allowed to stand for 2 h in...

Embodiment 3

[0070] Example 3 Preparation of Anti-colored clayromycin monoclonal antibody

[0071] Anti-colored clayromycin monoclonal antibodies were produced by the preservation of hybridoma cell line ST03, which was previously numbered CCTCN No. C2013187, which is specifically prepared in advance according to the method reported in the patent No. 201410115952.8. The specific preparation method is as follows: BALB / C mice are treated in advance, and the hybridoma cell line ST03 is injected into the abdomen of the mouse. After about one week, the ascites are collected, and the anti-sulfate method is purified. Monochromycin monoclonal antibody. Specific purification steps: mouse deposit filtrate filter paper, 12000 r / min, 4 ° C, 15 min centrifugal collected supernatant, absorbed into a quadroyroid of acetate buffer, slowly add orthodaculipine while stirring while stirring. To the endic acid end volume of 30 to 35 μl / ml, stirred at room temperature for 30 to 60 min, and then allowed to sta...

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Abstract

The invention discloses a time-resolved fluorescent reagent kit for synchronous detection of fusarium diacetate, aflatoxin B1 and versicolor. Including immunochromatography time-resolved fluorescent test strips and sample reaction bottles containing europium-labeled anti-sarcienol diacetate, aflatoxin B1, and versicolor monoclonal antibodies, the immunochromatography time Distinguish one side of the fluorescent test strip from top to bottom to arrange water-absorbing pads, detection pads and sample pads in sequence, and adjacent pads overlap and connect at the joints. A quality control line and a detection line are arranged horizontally from top to bottom. The quality control line is coated with a rabbit anti-mouse polyclonal antibody, and the number of detection lines is three, which are respectively coated with each toxin-protein conjugate. It can realize the rapid and simultaneous detection of three mycotoxins, Fusarenol diacetate, Aflatoxin B1, and Aspergillus versicolor.

Description

Technical field [0001] The present invention provides a fungal toxin time resolution fluorescent immunogenic examination kit, specifically a synchronous detection of dicetate 镳 镰 烯 (serpentin), xenojamin B1, a time to distinguish of mixed contamination of varicolorin Fluorescent kit and preparation method. Background technique [0002] Diacetic acid squat sicklelene (4, 15-DiaceToxyscirpenol, DAS) is a monohydrate toxin produced by Squam, Mycobika, Wirkeese. DAS mainly pollutes cereals and feed, which is toxic to animal bone marrow, brain, heart, lymphatic, testicular, thymine, nerve cell, etc., can also cause gastrointestinalitis, antibody reduction, eye and body edema. The yellow bellisxin is a class of toxic secondary metabolites produced by crops such as Asperillus flavus and parasitus, asperg illusparasiticus, mainly including B1, B2, G1, G2, M1, M2. The toxicity of the yellowlaxin B1 is 10 times that of potassium cyanide, 68 times the cream, is listed as a class carcinogen ...

Claims

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Application Information

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Patent Type & Authority Patents(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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