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High-flux ultra-sensitive fluorescent gold nano-cluster immunochromatographic test strip and application thereof

A technology of fluorescent gold nanometers and test strips, which can be used in measuring devices, analytical materials, instruments, etc., and can solve the problem of low fluorescence quantum yield

Inactive Publication Date: 2020-03-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently widely used fluorescent gold nanoclusters are synthesized with bovine serum albumin as a ligand, and the fluorescence quantum yield is low (<10%), so it is difficult to use it as a marker in rapid immunoassay technology.

Method used

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  • High-flux ultra-sensitive fluorescent gold nano-cluster immunochromatographic test strip and application thereof
  • High-flux ultra-sensitive fluorescent gold nano-cluster immunochromatographic test strip and application thereof
  • High-flux ultra-sensitive fluorescent gold nano-cluster immunochromatographic test strip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1: Preparation of clenbuterol hydrochloride and ractopamine fluorescent gold nanocluster immunochromatographic test strips

[0078] 1. Preparation of fluorescent detection probes

[0079] 1. Preparation of fluorescent gold nanoclusters

[0080] The fluorescent gold nanocluster (Arg-ATT-AuNCs) of the present invention is a fluorescent nanoparticle synthesized by a two-step method with 6-aza-2-thymine as a ligand and arginine as a stiffener. The guanidine group in the oxidizing agent arginine forms a "host-guest recognition" effect with 6-aza-2-thiothymine through hydrogen bonding, and the carboxyl group of arginine is exposed to the outside, making it easy to modify. For the preparation method of fluorescent gold nanoclusters, please refer to the literature: Fabrication of Water-Soluble, Green-Emitting Gold Nanoclusters with a 65% Photoluminescence Quantum Yield via Host–Guest Recognition, Deng H H, Shi X Q, Wang F F, et al.Chemistry of Materials, 2017, method ...

Embodiment 2

[0102] Example 2: Qualitative and quantitative detection methods of clenbuterol hydrochloride and ractopamine in pig urine samples

[0103] 1. Qualitative testing

[0104] Add 100 μl of the sample to be tested (pig urine sample tested negative for both clenbuterol hydrochloride and ractopamine) into the microwell containing 3 μl of the fluorescent detection probe solution prepared in step 1 of Example 1, mix and incubate for 3 min, Then add the incubated solution dropwise on the sample pad of the test strip, and after fifteen minutes, observe the color development of the detection line and the quality control line of the test strip under an ultraviolet light.

[0105] If the T1 line is not colored, the T2 line is not colored, and the C line is green, then the sample to be tested contains clenbuterol hydrochloride and ractopamine;

[0106] If the T1 line shows no color, the T2 line shows green, and the C line shows green, then the sample to be tested contains clenbuterol hydro...

Embodiment 3

[0116] Example 3. Application of Fluorescent Gold Nanocluster Immunochromatography

[0117] 1. Preparation of samples to be tested

[0118] The standard solutions 5, 7 and 8 in Example 2 were used as samples to be tested.

[0119] 2. Detection of clenbuterol hydrochloride and ractopamine

[0120] Add the sample to be tested into the microwell containing 3 μl fluorescent detection probe solution, mix and incubate for 3 minutes, then add the incubated solution dropwise on the sample pad of the test strip, and read the test strip with fluorescence after 15 minutes An instrument (ESE-Quant LR3, QIAGEN Co., Ltd., Germany) reads the fluorescence signals on the detection line and the quality control line of the test strip, and obtains the fluorescence intensities of T1, T2 and C lines, respectively. substitute it into Figure 4 Calculate the standard curve to obtain the content of clenbuterol hydrochloride and ractopamine in the test sample and calculate the recovery rate and coef...

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Abstract

The invention discloses a high-flux ultra-sensitive fluorescent gold nano-cluster immunochromatographic test strip and application thereof. The fluorescent gold nano-cluster immunochromatography teststrip comprises a sample pad, a nitrocellulose membrane, absorbent paper and a PVC viscous bottom plate; the nitrocellulose membrane is coated with two target object detection lines and a quality control line. The invention also discloses a complete set of fluorescent gold nano-cluster coupling antibody probe, which is composed of a streptavidin fluorescent gold nano-cluster labeled biotinylated anti-small molecule compound A antibody and a streptavidin fluorescent gold nano-cluster labeled biotinylated anti-small molecule compound B antibody. The fluorescent gold nano-cluster is a green fluorescent gold nano-cluster which takes 6-aza-2-thiopyrimidine and arginine as ligands and has super-strong fluorescence. The test strip has the advantages that the method is simple and rapid, the resultis visual and accurate, quantitative / semi-quantitative detection can be achieved, and high-flux ultra-sensitive online detection of veterinary drug residues can be achieved.

Description

technical field [0001] The invention belongs to the field of veterinary drug detection in food safety, and in particular relates to a high-throughput ultrasensitive fluorescent gold nano-cluster immunochromatographic test strip and an application thereof. Background technique [0002] Immunochromatography technology is a unique immune analysis method that appeared in the early 1980s. It has the advantages of fast, simple, sensitive and convenient, and low price. It can realize online rapid detection and monitoring of target objects. It is currently used in food safety and medical diagnosis , environmental monitoring and other fields have been widely used. The immunochromatographic technique based on colloidal gold as a marker is the most successful and widely used rapid screening method, but its sensitivity is often difficult to meet people's requirements. Therefore, it is necessary to establish a high-throughput and ultra-sensitive immunochromatographic method for the simu...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/533
CPCG01N33/533G01N33/577
Inventor 江海洋沈建忠王战辉温凯彭涛郑丕苗
Owner CHINA AGRI UNIV
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