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Fluorescence immunochromatography test paper for detecting ochratoxin A

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

Inactive Publication Date: 2016-09-28
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 immunochromatography test paper for detecting ochratoxin A
  • Fluorescence immunochromatography test paper for detecting ochratoxin A
  • Fluorescence immunochromatography test paper for detecting ochratoxin A

Examples

Experimental program
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Effect test

Embodiment 1

[0061]The preparation of fluorescent immunochromatographic test paper for detecting ochratoxin A mainly includes: preparation of OTA artificial antigen, preparation of OTA monoclonal antibody or polyclonal antibody, preparation of OTA antibody labeled with graphene oxide fluorescent nanomaterials, cellulose film layer Preparation and assembly of immunochromatographic test paper.

[0062] 1. Preparation of coupled OTA carrier protein

[0063] The active ester method is used to couple the OTA with the carrier protein to prepare the artificial antigen.

[0064] The specific steps are: accurately weigh 1.3mg OTA, 2mg dicyclohexylcarbodiimide (DCC) and 1.2mg N-hydroxysuccinimide (NHS) and dissolve them in 1.0mL methanol solution successively, and stir for 2 hours at room temperature in the dark , carry out a sufficient chemical reaction, and name it A liquid. Weigh 5mg of bovine serum albumin (BSA) and dissolve it in 2mL of PBS (0.01M, pH7.4) buffer solution, named solution B. U...

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 OTA monoclonal antibody or polyclonal antibody.

[0093] The NaYF 4 : Yb, the preparation method of the OTA monoclonal antibody or 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 NH 4 F solution 7mL, under the condition of 25 ℃ dark stirring 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

[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 OTA monoclonal or polyclonal antibody.

[0107] The NaGd (WO 4 ) 2 : Eu 3+ The preparation method of the OTA 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 fluorescence immunochromatography test paper for detecting ochratoxin A. The fluorescence immunochromatography test paper comprises a supporting body and an absorbing layer fixed to the supporting body, and the absorbing layer sequentially comprises an absorbing fiber layer, a fluorescent antibody fiber layer, a cellulose membrane layer and a water absorbing material layer from the test end; invisible detection imprinting printed with a coupling OTA carrier protein solution and invisible detection imprinting printed with a goat-anti-mouse IgG, rabbit-anti-mouse IgG and goat-anti-rabbit IgG antibody solution are arranged on the cellulose membrane layer; the fluorescent antibody fiber layer is made of a glass fiber cotton adsorbing fluorescent antibody, and the fluorescent antibody is an OTA monoclonal antibody or polyclonal antibody marked with an oxidized graphene fluorescence nanometer material, NaYF4:Yb, Tm nanometer particles or NaGd(WO4)2:Eu3+ nanometer particles. The test paper strip has the advantages of being high in specificity, sensitivity and stability, good in safety, convenient and fast. On-site quantitative detection can be achieved under a portable fluorescence reading instrument, and requirements of people at different levels can be met.

Description

technical field [0001] The invention relates to an immunochromatographic test paper, in particular to a fluorescent immunochromatographic test paper for detecting ochratoxin A. Background technique [0002] Ochratoxin is another mycotoxin that has attracted worldwide attention after aflatoxin. It is a group of important food-contaminating mycotoxins produced by 7 species of Aspergillus in the genus Aspergillus and 6 species in Penicillium in the genus Penicillium. Ochratoxin A (OTA) has the highest amount, the most serious pollution to agricultural products, and the most closely related to human health. OTA mainly poisons the kidney and liver of animals, and the kidney is the first target organ, which has strong teratogenic, carcinogenic and mutagenic effects. my country's GB2761-2011 national food safety standard stipulates that the OTA limit standard in beans and their products, grains and their rolled products is 5 μg / kg; on July 5, 2012, the European Commission issued ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
CPCG01N33/558G01N2333/38G01N2333/385
Inventor 李靖靖职爱民闵玉涛程丽英郭林王岚张小梅聂卉赵国欣张晓宇杨联芝
Owner ZHONGZHOU UNIV
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