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Clenbuterol detection test paper and clenbuterol detection method based on fluorescence immunochromatography technology

A fluorescence immunochromatography, detection test paper technology, applied in biological testing, measuring devices, analysis materials, etc., can solve the problems of easy false positive results, inability to quantitative detection, long reaction time, etc., and achieve simple and fast preprocessing. The effect of ultra-high detection accuracy and sensitivity, and short processing time

Pending Publication Date: 2020-12-25
JIAXING CHAOYUNFAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA has the advantages of simple operation, relatively low detection cost, and relatively high sensitivity, but it requires multi-step washing, and the reaction time is long, and its main detection samples are urine and plasma, and false positive results are prone to occur in the detection of animal tissue samples
GLFIS has the advantages of simple operation, rapidity, and low cost, but the sensitivity of this method is relatively low, and it cannot be quantitatively detected

Method used

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  • Clenbuterol detection test paper and clenbuterol detection method based on fluorescence immunochromatography technology
  • Clenbuterol detection test paper and clenbuterol detection method based on fluorescence immunochromatography technology
  • Clenbuterol detection test paper and clenbuterol detection method based on fluorescence immunochromatography technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038]Such asfigure 1 As shown, the clenbuterol test paper based on the fluorescence immunochromatography technology of the present invention includes a support base 5 and a sample pad 1, a marker pad 2, a capture film 3, and a water-absorbing pad 4 attached to the upper surface of the support base 5, adjacent to each other. Each part of is overlapped at the edge joints, the capture film 3 contains C line 6 and T line 7; the label pad 2 contains clenbuterol detection antibody labeled with fluorescent microspheres or contains fluorescent microspheres Labeled control protein or antibody; the T line 7 on the capture membrane 3 contains clenbuterol antigen; the C line 6 contains a secondary antibody that can bind to the clenbuterol detection antibody or contains a protein or antibody that can be used as a positive control.

[0039]In this embodiment, the fluorescent microsphere-labeled control protein or antibody contained in the marker pad 2 can be streptavidin or rabbit antibody; a T lin...

Embodiment 2

[0043]based onfigure 1 The present invention provides a preparation method for the clenbuterol test paper shown, which specifically includes the following steps:

[0044]S1: Prepare fluorescent microspheres and clenbuterol antigen antibodies;

[0045]S2: Fluorescent microspheres are coupled with clenbuterol antibodies to obtain clenbuterol antibodies labeled with fluorescent microspheres;

[0046](201) Mix 200μL of 20mM PBS, pH7.4, 1% solid content fluorescent microspheres with 6mg carbodiimide, shake the reaction at room temperature for 30min, centrifuge at 12000rpm for 10min, wash twice with borate buffer to obtain activation Fluorescent microspheres behind;

[0047](202) Resuspend the activated fluorescent microspheres in 300μL borate buffer, add 200μL of 1mg / mL clenbuterol antibody to the microsphere suspension, and shake and react for 2.5h at room temperature;

[0048](203) Centrifuge the reacted solution at 12000 rpm for 10 minutes, add 400 μL of 0.25M ethanolamine and shake for another 30 m...

Embodiment 3

[0060]In this example, clenbuterol hydrochloride was used as the clenbuterol antigen and antibody to prepare test strips according to the steps in Example 2, and the prepared test strips were used to detect clenbuterol hydrochloride in animal tissue samples The specific steps are as follows:

[0061]S1: 100μL of clenbuterol hydrochloride solution containing 0μg / L, 0.5μg / L, 1μg / L, 2μg / L, 10μg / L, 20μg / L, 50μg / L, 100μg / L was added dropwise to the immune layer. Analyze the sample pad of the test paper, and detect the fluorescence signal with a fluorescence detector after reacting for 15 minutes;

[0062]S2: Then plot the fluorescence intensity vs. clenbuterol concentration to obtain a standard curve of fluorescence intensity vs. clenbuterol concentration;

[0063]S3: Treat the animal tissue sample with 0.5%-10% NaCL for 1min-20min to obtain a sample solution;

[0064]S4: Dilute the obtained sample solution 2-10 times with the diluent, take 100 μL of the sample solution containing clenbuterol hydroc...

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Abstract

The invention discloses a clenbuterol detection test paper and detection method based on a fluorescence immunochromatography technology. The method comprises the following steps of preparing and assembling test paper, and using immunochromatographic test paper to perform quantitative detection of clenbuterol. The detection test paper comprises a supporting bottom plate, a sample pad, a marker pad,a capture film and a water absorption pad; the sample pad, the marker pad, the capture film and the water absorption pad are adhered to the upper surface of the supporting bottom plate; the capture film contains a C line and a T line; the marker pad contains a clenbuterol detection antibody marked by fluorescent microsphere or contains a control protein or antibody marked by the fluorescent microsphere; the T line on the capture film contains a clenbuterol antigen; and the C line contains a secondary antibody capable of being combined with the clenbuterol detection antibody or contains a protein or antibody capable of serving as a positive control. The fluorescent microsphere is coupled with the clenbuterol antibody to obtain the clenbuterol antibody marked by the fluorescent microsphere,so that the prepared test paper has higher sensitivity in the detection process, and meanwhile, the reliability of the method for quantitatively detecting clenbuterol component through fluorescence immunochromatography is also guaranteed.

Description

Technical field[0001]The present invention relates to the field of biological immune detection, covering nano material technology, immunology technology and lateral flow chromatography technology. More specifically, the present invention relates to a clenbuterol detection test paper and method based on fluorescence immunochromatography technology.Background technique[0002]Clenbuterol is a hormone drug that promotes the growth of lean meat and inhibits the growth of fat. The basic chemical structure of this class of drugs is β-phenethylamine, therefore, it is also called β-agonist (β-agonist). According to its substituent type, it can be divided into three types: aniline, phenol, and benzenediol. Big category. Representative drugs are clenbuterol hydrochloride and ractopamine. Early people discovered that adding clenbuterol into animal feed can promote protein synthesis in animals, inhibit fat synthesis and accumulation, and increase lean meat rate, so it is widely used in aquacultur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/558G01N33/533
CPCG01N33/94G01N33/582G01N33/585G01N33/558G01N33/533
Inventor 杨挥
Owner JIAXING CHAOYUNFAN BIOTECH
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