Fluorescent microsphere-colloidal gold dual-color qualitative and 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 measuring devices, analytical materials, instruments, etc., can solve the problems of large sample background interference, no longer accurate and reliable results, and long time-consuming, so as to eliminate false positive results , Realize the result data, avoid the effect of matrix interference

Active Publication Date: 2020-11-03
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 dual-color qualitative and quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof
  • Fluorescent microsphere-colloidal gold dual-color qualitative and quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof
  • Fluorescent microsphere-colloidal gold dual-color qualitative and quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Fluorescence microsphere-colloidal gold double color qualitative and quantitative immunochromatographic test strip preparation for detecting clenbuterol hydrochloride (optimum ratio of fluorescent microsphere and colloidal gold antibody labeling complex)

[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 the diazo method, the coupling protein is bovine serum albumin, and the coupling ratio is 1:10-1:100. After coupling, the CLE-BSA is obtained by dialysis and purification.

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

[0061] CLE-BSA conjugate and goat anti-mouse antibody are coated on nitrocellulose membrane: Dilute the CLE-BSA conjugate with 0.05M pH 7.2 PBS (phosphate buffered saline) to make the concentration 0.2mg / mL, The resulting solution was sprayed on the membra...

Embodiment 2

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

[0080] The difference from 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, and the amount of colloidal gold antibody is accurate to 30ng / bar, fluorescent microsphere The antibody amount was 6 ng / strip marker concentration to spray the binding pad. Spray on 30×0.8cm glass fiber membrane, vacuum dry at 25°C for 1 to 2 hours, and place in a drying cabinet for later use. The quantitative detection results of fluorescent microspheres are: the concentration of the standard curve under this condition is: 0, 0.1, 0.3, 0.9, 1.5, 2.7 ng / mL, R 2 0.9058, IC 5...

Embodiment 3

[0085] Example 3: Preparation of fluorescent microsphere-colloidal gold double color qualitative and quantitative immunochromatographic test strip for detecting clenbuterol hydrochloride (the proportion of colloidal gold labeled complexes increases)

[0086] The difference from Example 1 is:

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

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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 veterinary drug residue detection in food safety, and specifically relates to a time-resolved fluorescent microsphere-colloidal gold double color qualitative and quantitative immunochromatographic test strip that can simultaneously qualitatively and quantitatively detect clenbuterol in a sample and a preparation method thereof. Background technique [0002] Clenbuterol Hydrochloride (CLE) is a synthetic β-adrenergic stimulant, which has the effect of expanding the bronchus. It is commonly used to prevent and treat lung diseases such as asthma and emphysema. But when its applied dose reaches 5-10 times the therapeutic dose, it can increase muscle synthesis and reduce fat deposition, so it is commonly called "Clenbuterol". In animal husbandry production, some illegal users add it as a growth promoter to animal feed to increase the growth rate of lean meat and also cause clenbuterol residues in animal food. When people ea...

Claims

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Application Information

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Patent Type & Authority Patents(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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