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Fluorescent immunochromatography test paper for detecting aflatoxin B1

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

Inactive Publication Date: 2016-10-12
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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  • Fluorescent immunochromatography test paper for detecting aflatoxin B1
  • Fluorescent immunochromatography test paper for detecting aflatoxin B1
  • Fluorescent immunochromatography test paper for detecting aflatoxin B1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Detection of Aflatoxin B 1 The preparation of fluorescent immunochromatographic test paper mainly includes: AFB 1 Preparation of artificial antigen, AFB 1 Preparation of monoclonal antibody or polyclonal antibody, graphene oxide fluorescent nanomaterial labeling AFB 1 Preparation of antibody, preparation of cellulose membrane layer and assembly of immunochromatography test paper and other steps.

[0061] 1. Coupled with AFB 1 Preparation of carrier protein

[0062] will AFB 1 Coupling with carrier protein to prepare artificially synthesized antigen.

[0063] Weigh 10mg of bovine serum albumin (BSA) and place it in a 10mL screw bottle, add 0.13mol / L NaHCO 3 The solution was made into a 10% BSA activation solution with a mass fraction of 10%, and the pH was adjusted to 7.6. Weigh 1mg AFB 1 , 1.073mg N,N-dicyclohexylcarboimide and 0.598mg N-hydroxysuccinimide (NHS), dissolved in anhydrous tetrahydrofuran, shaken at 30°C for 4h, then centrifuged at 4000r / min for 15m...

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 : AFB labeled with Yb,Tm nanoparticles 1 Monoclonal or polyclonal antibodies.

[0092] The NaYF 4 : AFB labeled with Yb,Tm nanoparticles 1 A method for preparing a monoclonal antibody or a polyclonal antibody, comprising 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 4 F 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...

Embodiment 3

[0105] 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 AFB 1 Monoclonal or polyclonal antibodies.

[0106] The NaGd (WO 4 ) 2 : Eu 3+ Nanoparticle-labeled AFB 1 A method for preparing a monoclonal antibody or a polyclonal antibody, comprising the following steps:

[0107] (1) NaGd (WO) was prepared by hydrothermal synthesis 4 ) 2 : Eu 3+ nanoparticles

[0108] 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...

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Abstract

The invention discloses fluorescent immunochromatography test paper for detecting aflatoxin B1. The fluorescent immunochromatography test paper comprises a supporting body and an adsorption layer fixed to the supporting body, and 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 testing end; the cellulose membrane layer is provided with invisible detection imprints imprinted by an AFB1-coupled carrier protein solution and invisible control imprints imprinted by a goat anti-mouse IgG antibody solution or a rabbit anti-mouse IgG antibody solution or a goat anti-rabbit IgG antibody solution; the fluorescent antibody fiber layer is made of glass fiber cotton adsorbing fluorescent antibodies, wherein the fluorescent antibodies are AFB1 monoclonal antibodies or AFB1 polyclonal antibodies which are labeled with graphene oxide fluorescent nanometer materials or NaYF4:Yb, Tm nanoparticles or NaGd(WO4)2:Eu<3+> nanoparticles. The test paper strip has the advantages of being high in specificity, sensitivity and stability, good in safety and easy, convenient and rapid to manufacture, and can achieve on-site quantitative detection in the presence of a portable fluorescent reading meter and meet the requirements of personnel at different levels.

Description

technical field [0001] The invention relates to an immunochromatographic test paper, in particular to a method for detecting aflatoxin B 1 fluorescent immunochromatographic test strips. Background technique [0002] Aflatoxin B 1 (Aflatoxin B 1 , called AFB 1 ) are fungal secondary metabolites produced by Aspergillus flavus, Aspergillus parasiticus and Aspergillus nomius. AFB 1 It often naturally exists in crops such as peanuts, cottonseed, corn, wheat and rice, and has a high detection rate. It is highly toxic and carcinogenic, and belongs to Class I carcinogens. my country AFB 1 There are many reports on the hazards of food and feed, and the detection rate is as high as 80% to 100%. It can inhibit animal immune function, reduce animal production performance, cause secondary infection, and even death of animals. At the same time, it can remain in animal products and cause Threat to human health. Therefore for AFB 1 It is meaningful to carry out effective rapid dete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/533G01N33/543
CPCG01N33/533G01N33/54346G01N33/558
Inventor 李靖靖职爱民聂卉赵国欣王岚闵玉涛张小梅孙浩冉郭林杨联芝
Owner ZHONGZHOU UNIV
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