Fluorescence immunoassay chromatography test strip for detecting vomitoxin

A technique of fluorescence immunochromatography and vomitoxin, applied in fluorescence/phosphorescence, material analysis through optical means, measuring devices, etc., can solve problems such as insufficient luminous efficiency and few types of fluorescent materials

Inactive Publication Date: 2016-08-17
ZHONGZHOU UNIV
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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 immunoassay chromatography test strip for detecting vomitoxin
  • Fluorescence immunoassay chromatography test strip for detecting vomitoxin
  • Fluorescence immunoassay chromatography test strip for detecting vomitoxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061]The preparation of fluorescent immunochromatographic test paper for detecting vomitoxin mainly includes: the preparation of DON artificial antigen, the preparation of DON monoclonal antibody or polyclonal antibody, the preparation of DON antibody labeled with graphene oxide fluorescent nanomaterials, and the preparation of cellulose film layer And steps such as the assembly of immunochromatographic test paper.

[0062] 1. Preparation of coupled DON carrier protein

[0063] Coupling DON with carrier protein to prepare synthetic antigen

[0064] Weigh 1.5 mg of dicyclohexylcarbodiimide and 2.7 mg of N-hydroxysuccinimide (NHS), and dissolve them in 200 μL of N,N-dimethylformamide to obtain liquid A and liquid B. Weigh 4 mg of bovine serum albumin (BSA) and dissolve it in 200 μL of 0.01 mol / L, pH 7.4 PBS to obtain solution C, which is stored in the refrigerator until use. 2.0 mg DON was dissolved in 350 μL of DMF to obtain solution D. Add liquid A and liquid B to liquid D...

Embodiment 2

[0092] 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 DON monoclonal antibody or polyclonal antibody.

[0093] The NaYF 4 : Yb, the preparation method of the DON monoclonal antibody or the polyclonal antibody of Tm nanoparticle labeling, comprises the following steps:

[0094] (1) Preparation of NaYF by hydrothermal synthesis 4 :Yb,Tm nanoparticles:

[0095] 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 add 0.4mol / L sodium citrate solution 2mL, fully react for 1h at 25°C in the dark; add 1.5mol / L NH4 F 7mL, stirred for 0.5h in the dark at 25°C, 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 double distilled water, and dr...

Embodiment 3

[0106] 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 DON monoclonal or polyclonal antibody.

[0107] The NaGd (WO 4 ) 2 : Eu 3+ The preparation method of the DON monoclonal antibody or polyclonal antibody of nanoparticle labeling, comprises the following steps:

[0108] (1) NaGd (WO) was prepared by hydrothermal synthesis 4 ) 2 : Eu 3+ nanoparticles

[0109] 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 immunoassay chromatography test strip for detecting vomitoxin. The fluorescence immunoassay chromatography test strip comprises a supporting body and an adsorption layer fixed to the supporting body. The adsorption layer sequentially comprises an adsorption fiber layer, a fluorescent antibody fiber layer, a cellulose membrane layer and a water absorption material layer starting from the test end. Invisible detection prints printed by a coupling DON carrier protein solution and invisible contrast prints printed by a goat anti-mouse IgG or rabbit anti-mouse IgG or goat anti-rabbit IgG antibody solution are arranged on the cellulose membrane layer. The fluorescent antibody fiber layer is made from glass fiber cotton for adsorbing fluorescent antibodies. The fluorescent antibodies are graphene oxide fluorescent nanometer materials or NaYF4:Yb or Tm nano-particles or DON monoclonal antibodies or polyclonal antibodies marked by NaGd(WO4)2:Eu3+ nano-particles. The test strip has the advantages of being high in specificity, sensitivity and stability, good in safety and easy, convenient and rapid to use, can achieve on-site quantitative detection under a portable fluorescent reading device and can meet the requirements of people of different levels.

Description

technical field [0001] The invention relates to an immunochromatographic test paper, in particular to a fluorescent immunochromatographic test paper for detecting vomitoxin. Background technique [0002] Vomitoxin (Vomitoxin, called DON) is mainly a toxin produced by Fusarium graminearum, Fusarium oxysporum, Fusarium pink, Fusarium moniliforme, Fusarium brick red, Fusarium nivalum, etc. The toxin is polluted wheat, The most widespread and common mycotoxins in oats, corn, barley and other grains (feeds) and their products. At present, food and feed all over the world are polluted by it, seriously endangering human and animal health. [0003] DON mainly acts on cells and tissues with active proliferation, such as lymphocytes, mucosal epithelium, bone marrow and liver. A variety of enzyme activities have obvious effects, which can cause lipid peroxidation damage in animals. At the third conference on additives and pollutants jointly held by the Food and Agriculture Organizat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N21/64
CPCG01N21/6486G01N33/558G01N33/577
Inventor 职爱民聂卉李建新闵玉涛程丽英张小梅孙浩冉郭林杨联芝李靖靖
Owner ZHONGZHOU UNIV
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