Chemiluminiscence immunodetection reagent kit for detecting salbutamol

A chemiluminescent immunoassay and detection kit technology, which is applied in the field of immunological detection, can solve the problems of prolonging the detection time, and achieve the effects of increasing sensitivity, reducing incubation time, and simple instruments

Inactive Publication Date: 2013-02-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of the method used by Xu Chuanlai et al. is that the primary antibody and the secondary antibody need to be used at the same time (the secondary antibody is an enzyme-labeled antibody). After the addition of the two antibodies, incubation is required, which prolongs the detection time.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1) Preparation of each component of the kit

[0051] Preparation of salbutamol hapten: acidify salbutamol, react with sodium nitrite in a low-temperature environment without light at 4°C, and generate intermediates containing diazonium cations. The diazotized albuterol was used as a hapten for subsequent synthesis of immune antigens and coating antigens.

[0052] Synthesis of bovine serum albumin-salbutamol immune antigen: the immune antigen was obtained by coupling salbutamol and bovine serum albumin (BSA) by diazotization method.

[0053] Synthesis of human serum albumin-salbutamol-coated antigen: the immune antigen was obtained by coupling salbutamol and human serum albumin (HSA) by diazotization.

[0054] Preparation of albuterol-peroxidase-labeled antibody: inject the above-mentioned immune antigen into Balb / C mice, and make it produce antibody serum after several booster immunizations. Take out the splenocytes of the immunized mice, add them to the centrifuge tu...

Embodiment 2

[0082] The chemiluminescent immunoassay kit for detecting salbutamol includes the following components:

[0083] (1) 48-well opaque white ELISA plates (6 strips x 8 wells) are coated with salbutamol-mouse serum albumin cross-linked complex, and are vacuum-sealed with aluminum film.

[0084] (2) 5 bottles of albuterol standard solution, the concentrations are respectively:

[0085] 0μg / L, 0.04μg / L, 0.36μg / L, 3.24μg / L, 29.16μg / L

[0086] (3) Salbutamol-horseradish peroxidase labeled antibody solution.

[0087] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0088] (5) 50 times concentrated buffer. Diluted 50 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L Tris-HCl buffer, pH6.9, containing 0.1% (v / v) Tween-20.

Embodiment 3

[0090] The chemiluminescent immunoassay kit for detecting salbutamol includes the following components:

[0091] (1) 96-well opaque white ELISA plate (12 strips x 8 wells) is coated with albuterol-egg albumin cross-linked complex and vacuum-sealed with aluminum film.

[0092] (2) 6 bottles of albuterol standard solution, the concentrations are respectively:

[0093] 0μg / L, 0.05μg / L, 0.25μg / L, 1.25μg / L, 6.25μg / L, 31.25μg / L.

[0094] (3) Salbutamol-horseradish peroxidase labeled antibody solution.

[0095] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0096] (5) 10 times concentrated buffer. Diluted 10 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L glycine-HCl buffer, pH6.9, containing 0.02% (v / v) Tween-20.

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Abstract

The invention provides a chemiluminiscence immunodetection reagent kit for detecting salbutamol, and belongs to the field of immunological detection. The reagent kit comprises a non-transparent white elisa plate coated with a salbutamol-carrier protein crosslinking compound, a salbutamol standard substance, a salbutamol-peroxidase labeled antibody, a luminous substrate solution and a concentrated buffer solution. The salbutamol-carrier protein crosslinking compound is obtained by coupling the salbutamol and a carrier protein through a mixed anhydride method or direct protein activation method, and the concentrated buffer solution contains a Tween-20 buffer solution. The reagent kit has the advantages of rapidness, simplicity, convenience, high sensitivity, low detection cost, good repeatability, high throughput and the like, and can be used for detecting the residual amount of the salbutamol in animal urine, blood, tissues, viscera and other samples.

Description

[0001] This application is a divisional application of the Chinese invention patent application "chemiluminescent immunoassay kit for detecting albuterol", the application number is 200810203436.5, and the application date is November 27, 2008. technical field [0002] The invention relates to a chemiluminescent immunoassay kit for detecting albuterol, which is used for detecting the content or residual amount of albuterol in animal-derived food such as animal tissue, viscera, blood, urine, feed, and feed raw materials. It belongs to the field of immunological detection. Background technique [0003] Salbutamol (Salbutamol, SAL) is a β-agonist. β stimulant is a kind of nutrient redistribution agent, which is a kind of phenylethanolamine derivatives with structure and function similar to epinephrine and norepinephrine. It can improve the ratio of meat to fat in fatty animals and reduce ketone body fat Content, increase lean meat rate, and can accelerate animal growth, so it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 陈峰梁晓翠卢庆鋆徐晓婴卢永红
Owner SHANGHAI JIAO TONG UNIV
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