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Fluorescence detection card for aflatoxin B1, preparation method of fluorescence detection card and method for detecting aflatoxin B1 in grain and oil

A technology for aflatoxin and fluorescence detection, which is applied in biochemical equipment and methods, measuring devices, bioreactors/fermenters for specific purposes, etc., can solve problems such as false positives and inability to realize on-site rapid detection, and achieve emission peaks Narrow, short detection time, simple operation

Pending Publication Date: 2022-05-24
GUANGZHOU ANNUO FOOD SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection of aflatoxin by ELISA has the advantages of high sensitivity, good recovery rate, strong specificity, etc., and can be used for qualitative and quantitative analysis, and has been widely used. However, this method has false positives and requires skilled technical operators disadvantages, so field fast detection is still not possible

Method used

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  • Fluorescence detection card for aflatoxin B1, preparation method of fluorescence detection card and method for detecting aflatoxin B1 in grain and oil
  • Fluorescence detection card for aflatoxin B1, preparation method of fluorescence detection card and method for detecting aflatoxin B1 in grain and oil

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A fluorescence detection card for aflatoxin B1, comprising a shell and a bottom plate arranged inside the shell, the shell including an upper shell and a lower shell that are fastened to each other, and the surface of the bottom plate is sequentially provided with a water absorbing material, a chromatography Membrane and sample binding pad; wherein, the surface of the chromatography membrane is provided with a quality control line and a detection line in turn; the upper shell is provided with an observation window and a sample injection hole; the sample binding pad is arranged below the sample injection hole, and the quality control line and detection line are arranged Set below the observation window; wherein, the polyester film in the sample binding pad is embedded with fluorescent microsphere-labeled aflatoxin B1 monoclonal antibody conjugate, and the quality control line of the chromatography film is coated with aflatoxin B1 antigen, the detection line is coated with...

Embodiment 2

[0051] The structure and preparation method of the detection card of Example 2 are the same as those of Example 1. The difference between Example 2 and Example 1 is that: the sample of Example 2 is rice, 1 g of the sample is weighed into a centrifuge tube, 3 mL of extract is added and fully mixed, centrifuged at 4000 r / min for 5 min, and the supernatant is taken out for use.

[0052] Wherein, the preparation method of the aflatoxin B1 monoclonal antibody conjugate labeled with fluorescent microspheres is as follows: take 90 ul of fluorescent microspheres into a centrifuge tube, add 900 ul of ultrapure water, mix well, and disperse by ultrasonic for 2 minutes. Add 60 ul of N-hydroxysuccinimide (NHS) and 60 ul of carbodiimide (EDC) solution with a concentration of 60 mg / ml respectively, mix with a mixer for 20 minutes, centrifuge at 15,000 rpm for 30 minutes, and remove the supernatant Add 1 ml of ultrapure water to the solution, ultrasonicate for 10 minutes to obtain a microsph...

Embodiment 3

[0054] The structure and preparation method of the detection card of Example 3 are the same as those of Example 1. The difference between Example 3 and Example 1 is that the extract used in the method for detecting aflatoxin B1 in grain and oil includes an ultrapure aqueous solution of methanol with a concentration of 15%.

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Abstract

The invention discloses a fluorescence detection card for aflatoxin B1, a preparation method of the fluorescence detection card and a method for detecting the aflatoxin B1 in grain and oil. A fluorescent microsphere labeled aflatoxin B1 monoclonal antibody conjugate is embedded in a sample combination pad of the detection card, an aflatoxin B1 antigen is coated on a quality control line of a chromatography membrane, and a goat anti-mouse antibody is coated on a detection line. The preparation method of the detection card comprises the following steps: adding a fluorescent microsphere labeled aflatoxin B1 monoclonal antibody conjugate into the sample combination pad for embedding treatment to obtain the fluorescent detection card with high sensitivity. According to the method for detecting the aflatoxin B1 in the grain and oil, a sample pretreatment method is limited, a specific extracting solution is selected, the extraction rate of the aflatoxin B1 in the grain and oil can be increased, aflatoxin B1 residues in a sample can be effectively extracted, and therefore measured data are more accurate.

Description

technical field [0001] The invention relates to the technical field of detecting aflatoxin B1, in particular to a fluorescence detection card for aflatoxin B1 and a preparation method thereof, and a method for detecting aflatoxin B1 in grains and oils. Background technique [0002] Aflatoxins are mainly produced by the toxigenic Aspergillus flavus, Aspergillus parasiticus and Aspergillus. Other molds such as Fusarium, Penicillium, Padium, and Rhizopus can also produce aflatoxins. Aflatoxin-contaminated grains and agricultural products mainly include corn, peanuts, nut food, rice, milk and dairy products, meat and aquatic products. Among them, peanuts and corn are the most polluted agricultural products, which have brought great harm to people's health. [0003] Currently commonly used detection methods for aflatoxin B1 include thin layer chromatography (TLC), high performance liquid chromatography (HPLC), colloidal gold immunochromatography (GICT) and enzyme-linked immunos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12M1/00G01N33/569G01N33/558G01N33/577G01N33/543G01N33/533
CPCG01N33/56961G01N33/558G01N33/577G01N33/54313G01N33/533G01N2333/38
Inventor 范彬王军杨鹏博温俊梅黄凤
Owner GUANGZHOU ANNUO FOOD SCI & TECH CO LTD
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