Chemiluminescence immune detection reagent kit for detecting ractopamine

A technique of chemiluminescence immunity and ractopamine, applied in the field of immunological detection, can solve problems such as prolonging detection time, achieve the effects of increasing sensitivity, simple instrument, and ensuring long-term effectiveness

Inactive Publication Date: 2009-04-22
SHANGHAI JIAO TONG UNIV
View PDF0 Cites 10 Cited by
  • 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 (...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1) Preparation of each component of the kit

[0050] Preparation of ractopamine hapten: acidify ractopamine, react with sodium nitrite in a low-temperature environment without light at 4°C, and generate intermediates containing diazonium cations. The diazotized ractopamine was used as a hapten for subsequent synthesis of immune antigens and coating antigens.

[0051] Synthesis of bovine serum albumin-ractopamine immune antigen: the immune antigen was obtained by coupling ractopamine and bovine serum albumin (BSA) by diazotization method.

[0052] Synthesis of human serum albumin-ractopamine-coated antigen: the immunization antigen was obtained by coupling ractopamine and human serum albumin (HSA) by a diazotization method.

[0053] Preparation of ractopamine-peroxidase-labeled antibody: inject the above-mentioned immunization antigen into Balb / C mice, and make it produce antibody serum after several booster immunizations. Take out the splenocytes of the immunized mice...

Embodiment 2

[0081] The chemiluminescent immunoassay kit for detecting ractopamine includes the following components:

[0082] (1) 48-well opaque white ELISA plates (6 strips x 8 wells) are coated with ractopamine-mouse serum albumin cross-linked complex, and are vacuum-sealed with aluminum film.

[0083] (2) 5 bottles of ractopamine standard solution, the concentrations are respectively:

[0084] 0μg / L, 0.04μg / L, 0.36μg / L, 3.24μg / L, 29.16μg / L

[0085] (3) Ractopamine-horseradish peroxidase labeled antibody solution.

[0086] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0087] (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

[0089] The chemiluminescent immunoassay kit for detecting ractopamine includes the following components:

[0090] (1) 96-well opaque white ELISA plate (12 strips x 8 wells) coated with ractopamine-egg albumin cross-linked complex, and vacuum-sealed with aluminum film.

[0091] (2) 6 bottles of ractopamine standard solution, the concentrations are respectively:

[0092] 0μg / L, 0.05μg / L, 0.25μg / L, 1.25μg / L, 6.25μg / L, 31.25μg / L.

[0093] (3) Ractopamine-horseradish peroxidase labeled antibody solution.

[0094] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0095] (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.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a chemiluminescent immunoassay kit for detecting ractopamine, and the kit belongs to the field of immunoassay. The kit is composed of a non-transparent white enzyme-labeled plate which is coated by a ractopamine-carrier protein cross-linked complex, a ractopamine standard product, a ractopamine-peroxidase-labeled antibody, luminescent substrate solution and concentrated buffer solution. The ractopamine-carrier protein cross-linked complex is obtained by coupling the ractopamine and a carrier protein through the mixed-anhydride method or the direct protein activation method, and the concentrated buffer solution contains Tween-20 buffer solution. The kit has the advantages of rapidness, simpleness, high sensitivity, low detection cost, good repeatability, high throughput and the like, and the kit can be used for detecting the residual ractopamine in animal urine, blood, tissues, visceral organs and other samples.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting ractopamine, which is used for detecting the content or residual amount of ractopamine in animal-derived foods such as animal tissue, viscera, blood, urine, feed, and feed raw materials. It belongs to the field of immunological detection. Background technique [0002] Ractopamine is a β-agonist. β-agonist is a kind of nutrient redistribution agent, which is a kind of phenylethanolamine derivatives similar in structure and function to adrenaline and norepinephrine. It can improve the ratio of meat to fat in fatty animals and reduce ketone bodies Fat content, increase lean meat rate, and can accelerate animal growth, so it is added to animal feed. Common β2-agonists include clenbuterol, ractopamine, and salbutamol. With the increasing supervision of clenbuterol (commonly known as "clenbuterol") in my country, the use of clenbuterol has gradually decreased, and the use of other β...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 陈峰卢庆鋆徐晓婴梁晓翠卢永红
Owner SHANGHAI JIAO TONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap