Immunofluorescence chromatography kit and method for simultaneous detection of aflatoxin b1 and zearalenone in vegetable oil

A technology of zearalenone and aflatoxin, which can be used in measurement devices, biological tests, analytical materials, etc., can solve the problems of complicated operation, long detection period, environmental factors and frequent operation effects of personnel.

Active Publication Date: 2021-07-06
广州敏捷生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing detection methods mainly include chromatography / mass spectrometry and immunoassay. The former belongs to physical and chemical detection methods. The pretreatment process is complicated, the degree of instrumentation is high, and the operation is cumbersome. long
Immunity methods currently mainly include ELISA and immunochromatography. The ELISA method has a long reaction process, frequent operations, and is easily affected by environmental factors and frequent operations by personnel. Accurate" and other aspects of the need
Colloidal gold and immunofluorescence chromatography are currently more popular in immunochromatography. Colloidal gold detection can be used for rapid qualitative detection, but the sensitivity is insufficient, and the accuracy of quantitative detection is still controversial.
The vegetable oil sample is viscous and inconvenient to absorb. If it is absorbed by the bottom, it is easy to absorb the upper layer of vegetable oil, and the process of removing impurities is not convenient.

Method used

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  • Immunofluorescence chromatography kit and method for simultaneous detection of aflatoxin b1 and zearalenone in vegetable oil
  • Immunofluorescence chromatography kit and method for simultaneous detection of aflatoxin b1 and zearalenone in vegetable oil
  • Immunofluorescence chromatography kit and method for simultaneous detection of aflatoxin b1 and zearalenone in vegetable oil

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Preparation of detection reagent card:

[0064] 1. The preparation of the sample pad is as follows:

[0065] (1) Sample pad treatment solution formula: 0.02M phosphate buffer solution with pH 7.4 containing 1% casein, 0.15M NaCl, 1.5% PEG 5000 and 1.5% TritonX-100;

[0066] (2) Apply the treatment solution prepared in the previous step evenly on the glass fiber RB65 at a concentration of 50 μl / cm2, and dry at 45°C for 16 hours.

[0067] 2. Preparation of the conjugation pad: Mix goat anti-chicken IgY polyclonal antibody-labeled microspheres, aflatoxin B1 detection microspheres and zearalenone detection microspheres in proportion, and spray evenly on the SB08 conjugation pad with an instrument, 45 °C for drying.

[0068] The preparation method of goat anti-chicken IgY polyclonal antibody-labeled microspheres is as follows:

[0069] (1) Add 1ml of 0.05M MES buffer solution to a 2ml centrifuge tube, then add 1mg of fluorescent microspheres, vortex and mix; add 4μl of 25...

Embodiment 2

[0104] Preparation of ID Chips Containing Brackets

[0105] Preparation of series standard products: Take 32μl aflatoxin B1 standard substance (2mg / kg) and 64μl zearalenone standard substance (100mg / kg) into 904μl methanol solution, mix well to get mixed standard substance 1 (aflatoxin B1 is 64 μg / kg, zearalenone is 6.4 mg / kg), and then diluted 5 times to obtain mixed standard 2, mixed standard 3, mixed standard 4, mixed standard 5, mixed standard 6 ;Take seven 15ml centrifuge tubes, add 2g blank vegetable oil (corn oil) sample without aflatoxin B1 and zearalenone to each tube, take 50μl mixed standard 1-6 and add it to 6 tubes, and add to the 7th tube 50μl methanol, mix well.

[0106] The final concentration gradient of aflatoxin B1 is: 1.6 μg / kg, 0.8 μg / kg, 0.4 μg / kg, 0.2 μg / kg, 0.1 μg / kg, 0.05 μg / kg, 0 μg / kg;

[0107] The final concentration gradient of zearalenone is: 160 μg / kg, 80 μg / kg, 40 μg / kg, 20 μg / kg, 10 μg / kg, 5 μg / kg, 0 μg / kg;

[0108] Add 2ml of saturated NaCl...

Embodiment 3

[0115] (1) Sample pretreatment and detection

[0116] Take 3 tubes of spiked vegetable oil (corn oil) and negative unspiked vegetable oil (corn oil), 2ml / tube, add 1ml of saturated saline and 2ml of acetonitrile to each tube, shake manually for 10s, let stand for stratification, and take the supernatant The solution was diluted 10 times with the diluent, to be tested;

[0117] (2) Detection

[0118] Take 100 μl of the solution to be tested and add it to the sample hole of the test card, and insert it into the instrument for reading after 10 minutes.

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Abstract

The invention discloses an immunoimmunofluorescence chromatographic kit and method for synchronously detecting the contents of aflatoxin B1 and zearalenone in vegetable oils, aiming to provide a high-sensitivity, strong anti-interference, which can simultaneously detect aflatoxin B1 in vegetable oils. Immunofluorescence chromatography kit for aspergillus toxin B1 and zearalenone content; its technical scheme is as follows: it includes a bottom plate, and the bottom plate is sequentially connected with an absorption pad, a reaction membrane, a binding pad, and a sample from left to right. Pad, the reaction membrane is marked with a detection line T1 containing aflatoxin B1 coupled antigen, a detection line T2 coupled with zearalenone antigen, and a line C containing chicken IgY quality control; the binding pad is coated The invention relates to a mixture containing aflatoxin B1 detection microspheres, zearalenone detection microspheres and goat anti-chicken IgY polyclonal antibody labeled microspheres, belonging to the technical field of biological detection.

Description

technical field [0001] The invention discloses an immunofluorescence chromatography kit, specifically, an immunofluorescence chromatography kit for synchronously detecting the contents of aflatoxin B1 and zearalenone in vegetable oil, and the invention also relates to Aspergillus flavus in vegetable oil Detection method of toxin B1 and zearalenone content. Background technique [0002] The food safety problem in China has always threatened the life and health of Chinese people. The recent "sour soup" poisoning incident is not too long ago. Nine people were poisoned and none survived. Mycotoxins in edible oils sometimes exceed the standard. Aflatoxin B1 and zearalenone in edible oils are commonly found in a variety of vegetable oils. There are various detection methods and each method is very mature, but each has its own advantages and disadvantages. Existing detection methods mainly include chromatography / mass spectrometry and immunoassay. The former belongs to physical and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/543G01N33/533G01N33/53G01N33/577
Inventor 汤永平刘卫林志欣羊晓珊梁展鹏高成李立
Owner 广州敏捷生物技术有限公司
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