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Fluorescent microsphere-colloidal gold double color development qualitative quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof

A technology of clenbuterol hydrochloride and immunochromatographic test paper, which is applied to measurement devices, analytical materials, instruments, etc., can solve the problems of long time consumption, large sample background interference, narrow detection linear range, etc., to avoid matrix interference, Realize the effect of digitizing results and eliminating false positive results

Active Publication Date: 2019-01-01
江西中德生物工程股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] ⑴General test strips are qualitatively analyzed by naked eye observation results, and accurate quantitative detection cannot be achieved;
[0006] (2) The matrix effect of different materials is obvious, and the sample background interference is large, which is easy to produce false positive results;
[0007] (3) The positive result of the test cannot be saved, usually after the judgment time is exceeded, the result is no longer accurate and reliable
[0009] ⑴In the actual test, the proportion of negative results is very high, but all test strips of this method need to be read by an instrument, otherwise the results cannot be obtained, and it takes a long time to test a large number of samples!
[0010] ⑵In quantitative detection, the detection department still needs a threshold to distinguish negative and positive, so the amount of data processing is relatively large;
[0011] (3) The sensitivity of a single fluorescent microsphere test strip is high, but in practical applications, the detection linear range of the higher sensitivity is narrower, and the quantitative concentration range of the sample is not suitable for the actual detection requirements

Method used

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  • Fluorescent microsphere-colloidal gold double color development qualitative quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof
  • Fluorescent microsphere-colloidal gold double color development qualitative quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof
  • Fluorescent microsphere-colloidal gold double color development qualitative quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Preparation of fluorescent microspheres-colloidal gold dual-color qualitative and quantitative immunochromatographic test strips for detection of clenbuterol hydrochloride (optimal ratio of fluorescent microspheres and colloidal gold antibody-labeled complexes)

[0056] 1. Preparation process of immunochromatographic test strips

[0057] 1. Preparation of Nitrocellulose Membrane

[0058] (1) Preparation of clenbuterol hydrochloride artificial antigen (CLE-BSA)

[0059] The coupling method is diazo method, the coupling protein is bovine serum albumin, and the coupling ratio is 1:10-1:100. After coupling, it is purified by dialysis to obtain CLE-BSA.

[0060] (2) Preparation of detection line and quality control line

[0061] CLE-BSA conjugates and goat anti-mouse antibodies were coated onto nitrocellulose membranes: dilute the CLE-BSA conjugates with 0.05M pH 7.2 PBS (phosphate buffered saline) to make the concentration 0.2mg / mL, The resulting solution was ...

Embodiment 2

[0079] Example 2: Preparation of fluorescent microsphere-colloidal gold dual-color qualitative and quantitative immunochromatographic test strips for detection of clenbuterol hydrochloride (the proportion of fluorescent microsphere-labeled complexes increases)

[0080] The difference with Example 1 is:

[0081] (1) After the preparation of fluorescent microsphere antibody complex and colloidal gold antibody complex is completed, calculate according to the amount of antibody on different markers in each test strip. The amount of antibody was sprayed on the conjugate pad at a marker concentration of 6 ng / strip. Spray it onto a 30×0.8cm glass fiber membrane, dry it in vacuum at 25°C for 1-2 hours, and put it in a drying cabinet for later use. The quantitative detection results of fluorescent microspheres are: under this condition, the concentration of the standard curve is: 0, 0.1, 0.3, 0.9, 1.5, 2.7ng / mL, R 2 0.9058, IC 50 0.33ng / mL, poor linearity. Colloidal gold visual ins...

Embodiment 3

[0085] Example 3: Preparation of fluorescent microspheres-colloidal gold dual-color qualitative and quantitative immunochromatographic test strips for detection of clenbuterol hydrochloride (the proportion of colloidal gold-labeled complexes increases)

[0086] The difference with Example 1 is:

[0087] (1) After the preparation of fluorescent microsphere antibody complex and colloidal gold antibody complex is completed, calculate according to the amount of antibody on different markers in each test strip, and the accurate amount of colloidal gold antibody is 50ng / strip, The amount of antibody was sprayed on the conjugate pad at a marker concentration of 3ng / strip. Spray it onto a 30×0.8cm glass fiber membrane, dry it in vacuum at 25°C for 1-2 hours, and put it in a drying cabinet for later use. The quantitative detection results of fluorescent microspheres are: under this condition, the concentration of the standard curve is: 0, 0.1, 0.3, 0.9, 1.5, 2.7ng / mL, R 2 0.9694, IC ...

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Abstract

The invention discloses a fluorescent microsphere-colloidal gold double color development qualitative quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and a preparation method thereof. A bottom plate is overlapped with a filter paper, a sample pad, a glass fiber mat sprayed with a fluorescent microsphere-clenbuterol hydrochloride monoclonal antibody complex anda colloidal gold-clenbuterol hydrochloride monoclonal antibody complex, a nitrocellulose membrane and an absorbent paper, and the nitrocellulose membrane is coated with a clenbuterol hydrochloride coupling antigen as a test line and coated with an anti-mouse antibody as a quality control line. If the color development of the detection line is lighter than that of the quality control line, the content of the clenbuterol hydrochloride in a sample is greater than 0.5 ng / mL, and qualitative determination is positive. The test strip qualitatively judged to be positive is directly inserted into a fluorescence reader, and the reader numerically quantifies a fluorescence signal of a fluorescent microsphere color development system to realize quantitative detection. The fluorescent microsphere-colloidal gold double color development qualitative quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and the preparation method thereof are mainly used for qualitative and quantitative detection of the clenbuterol hydrochloride in food safety testing.

Description

technical field [0001] The invention belongs to the field of detection of veterinary drug residues in food safety, and in particular relates to a time-resolved fluorescent microsphere-colloidal gold dual-color qualitative and quantitative immunochromatographic test strip capable of qualitatively and quantitatively detecting clenbuterol in a sample and a preparation method thereof. Background technique [0002] Clenbuterol Hydrochloride (CLE) is a synthetic β-adrenaline stimulant, which has the effect of dilating the bronchi, and is often used to prevent and treat lung diseases such as asthma and emphysema. But when its application dose reaches 5-10 times of the therapeutic amount, it can increase muscle synthesis and reduce fat deposition, so it is commonly known as "lean meat essence". In animal husbandry production, some illegal users add it as a growth promoter to animal feed to increase the growth ratio of lean meat, and also cause the residue of clenbuterol in animal fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/533
CPCG01N33/533G01N33/558G01N33/577
Inventor 陈媛赖卫华罗凯刘文娟伍燕华
Owner 江西中德生物工程股份有限公司
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