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Fluorescence immunochromatography test paper for detecting zearalenone

A technology of fluorescence immunochromatography and zearalenone, which is applied in biological testing, measuring devices, analysis materials, etc., can solve the problems of few types of fluorescent materials and insufficient luminous efficiency

Active Publication Date: 2016-11-02
JIAOZUO BAIAOTAIKE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation of up-conversion fluorescent materials and the research on test paper still have defects such as insufficient luminous efficiency and few types of fluorescent materials. The research and discussion on this aspect need to be strengthened

Method used

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  • Fluorescence immunochromatography test paper for detecting zearalenone
  • Fluorescence immunochromatography test paper for detecting zearalenone
  • Fluorescence immunochromatography test paper for detecting zearalenone

Examples

Experimental program
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Effect test

Embodiment 1

[0060]The preparation of fluorescent immunochromatographic test paper for detecting zearalenone mainly includes: preparation of ZEN artificial antigen, preparation of ZEN monoclonal antibody or polyclonal antibody, preparation of ZEN antibody labeled with graphene oxide fluorescent nanomaterials, cellulose membrane Layer preparation and assembly of immunochromatographic test paper.

[0061] 1. Preparation of coupled ZEN carrier protein

[0062] Coupling ZEN with carrier protein to prepare artificially synthesized antigen.

[0063] Weigh 10 mg of ZEN and dissolve it in 5 mL of pyridine, and stir for 12 h in the dark. Dry the resulting solution with nitrogen, add 5 mL of double-distilled water, extract 3 times with an equal volume of ethyl acetate, discard the water phase, and dry with nitrogen. Oil A was obtained. Weigh 2.3 mg of N-hydroxysuccinimide (NHS), 4.1 mg of dicyclohexylcarbodiimide (DCC) and 3.18 mg of A, mix thoroughly at room temperature overnight, a white precipi...

Embodiment 2

[0091] The preparation of the immunochromatographic test paper in embodiment 2 is basically the same as that in embodiment 1, the main difference being that the fluorescent antibody is NaYF 4 : Yb, Tm nanoparticles labeled ZEN monoclonal antibody or polyclonal antibody.

[0092] The NaYF 4 : Yb, the preparation method of the ZEN monoclonal antibody or polyclonal antibody of Tm nano particle label, comprises the following steps:

[0093] (1) Preparation of NaYF by hydrothermal synthesis 4 :Yb,Tm nanoparticles:

[0094] Take Y(NO 3 ) 3 Solution 5.5mL, Yb(NO 3 ) 3 Solution 0.5mL and Tm(NO 3 ) 3 Solution 0.5mL, mix well, then slowly add 0.4mol / L sodium citrate solution 2mL, fully react for 1h at 25°C in the dark; add 1.5mol / L NH 4F solution 7mL, stirred under the same conditions for 0.5h, with HNO 3 Adjust the pH value to 7.4, let it stand for 0.5h, add double distilled water to dilute to 30mL; then react at 220°C for 24h, cool to room temperature, filter, wash with doub...

Embodiment 3

[0104] The preparation of immunochromatography test paper in embodiment 3 is basically the same as embodiment 1, and the main difference is: the fluorescent antibody is NaGd (WO 4 ) 2 : Eu 3+ Nanoparticle-labeled ZEN monoclonal or polyclonal antibody.

[0105] The NaGd (WO 4 ) 2 : Eu 3+ The preparation method of the ZEN monoclonal antibody or polyclonal antibody of nanoparticle labeling, comprises the following steps:

[0106] (1) NaGd (WO) was prepared by hydrothermal synthesis 4 ) 2 : Eu 3+ nanoparticles

[0107] Weigh 7g Gd 2 o 3 and 7g Eu 2 o 3 , were added to 50mL of HNO 3 , heated to 80°C, kept warm and concentrated until all crystallized, and Gd(NO 3 ) 3 and Eu(NO 3 ) 3 ; 2 WO 4 2H 2 O was dissolved in deionized water and prepared as Na at a concentration of 0.2 mol / L 2 WO 4 solution; the prepared Gd(NO 3 ) 3 and Eu(NO 3 ) 3 Dissolved in deionized water to prepare Gd(NO 3 ) 3 solution and 15% Eu(NO 3 ) 3 solution, draw the prepared 5mL Gd(N...

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Abstract

The invention discloses a fluorescence immunochromatography test paper for detecting zearalenone. The test paper comprises a supporting body and an adsorption layer fixed on the supporting body, the adsorption layer sequentially comprises an adsorption fiber layer, a fluorescence antibody fiber layer, a cellulose film and a water absorbing material layer, and the cellulose film is provided with a ZEN-coupled carrier protein solution printed shealth detection blot and a goat anti-mouse IgGT, rabbit anti-mouse IgG or goat anti-rabbit IgG antibody solution printed shealth contrast blot; and the fluorescence antibody fiber layer is made of fluorescence antibody adsorbing glass fiber cotton, and the fluorescence antibody is a graphene oxide fluorescence nano-material, and a NaYF4:Yb, Tm nanoparticle or NaGd(WO4)2:Eu<3+> nano-particle labeled ZEN monoclonal antibody or polyclonal antibody. The test paper has the characteristics of strong specificity, high sensitivity, high stability, good safety, high simplicity, fastness, realization of onsite quantitative detection under a portable fluorescence reader, and meeting of needs of people with different levels.

Description

technical field [0001] The invention relates to an immunochromatographic test paper, in particular to a fluorescent immunochromatographic test paper for detecting zearalenone. Background technique [0002] Zearalenone (Zearalenone, called ZEN), also known as F-2 toxin, is a non-steroidal mycotoxin mainly produced by Fusarium pink and Fusarium graminearum. Widely present in moldy corn, sorghum, wheat, oats, barley and other cereal crops and milk, it is the most widely polluted Fusarium toxin in the world, and the detection rate of ZEN in feed raw materials and feeds in the country is as high as 100% . ZEN is a special toxic biotoxin with estrogen-like effects, which can cause abnormal reproductive functions such as abortion, stillbirth, and return to estrus in animals, and can also lead to growth decline, immunosuppression, infertility, deformity, etc. ZEN can also pass The food chain accumulates in the human body, which brings huge economic losses to the food industry, fee...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/52
CPCG01N33/525G01N33/558G01N33/577
Inventor 职爱民贾国超李镁娟张培蕾周志友赵强王自良宋莲军邱国庆
Owner JIAOZUO BAIAOTAIKE BIOTECH CO LTD
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