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Chemiluminescence immunodetection kit used for detecting phenylethanolamine A

A chemiluminescent immunoassay and detection kit technology, which is applied in the field of immunological detection, can solve the problems of low false positive rate of accurate detection, deviation of experimental results, expensive instruments, etc., to ensure long-term effectiveness, simple instruments, and increased sensitivity Effect

Inactive Publication Date: 2014-05-14
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography (HPLC) has the characteristics of high detection accuracy and low false positive rate; the main determination of high-performance liquid chromatography (HPLC) is that the instrument is expensive, the operation is cumbersome, time-consuming, and the detection cost is high
Gas chromatography-mass spectrometry (GC-MS) method has high sensitivity and low false positive rate; however, the sample of this method needs to be derivatized, which will cause deviations in the experimental results
Liquid chromatography-mass spectrometry (LC-MS) can detect urine, blood, hair and other samples, but similar to the above method, the experimental operation steps are cumbersome and the detection cost is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Preparation of each component of the kit

[0040] (1) Preparation of phenylethanolamine A hapten: acidify phenylethanolamine A, react with sodium nitrite in a low-temperature environment without light at 4° C. to generate an intermediate containing diazonium cations. Diazotized phenylethanolamine A is used as a hapten for subsequent synthesis of immune antigens and coating antigens;

[0041] (2) Preparation of phenylethanolamine A-bovine serum albumin (BSA) immunogen: coupling phenylethanolamine A with bovine serum albumin (BSA) by diazotization method to obtain immune antigen;

[0042] (3) Preparation of phenylethanolamine A-oval serum albumin (OVA) coated antigen: coupling phenylethanolamine A and egg serum albumin (OVA) by diazotization method to obtain coated antigen;

[0043](4) Preparation of phenylethanolamine A-peroxidase-labeled antibody: For 6-8 week-old female BALB / c mice (body weight 18-20 g), the high-dose immunization scheme is as follows: 160 μg pheny...

Embodiment 2

[0078] The chemiluminescent immunoassay kit for detecting phenylethanolamine A includes the following components:

[0079] (1) 96-well opaque white ELISA plate (8 wells x 12 strips) coated with phenylethanolamine A-mouse serum albumin conjugate, vacuum-sealed in an aluminum foil bag;

[0080] (2) 6 bottles of phenylethanolamine A standard solution, the concentrations are respectively:

[0081] 0 ng / mL, 0.05 ng / mL, 0.25 ng / mL, 1.25 ng / mL, 6.25 ng / mL, 31.25 ng / mL;

[0082] (3) phenylethanolamine A-horseradish peroxidase labeled antibody solution;

[0083] (4) Luminescence substrate A solution (luminol and enhancer), luminescence substrate B solution (urea hydrogen peroxide);

[0084] (5) 2 times concentrated sample diluent. Before use, please dilute 1:1 (1 part of concentrated diluent + 1 part of deionized water) to become the working sample diluent, and the diluted working sample diluent is 0.05 mol / L, pH 7.4 Tris-HCl buffer;

[0085] (6) 20 times concentrated was...

Embodiment 3

[0087] The chemiluminescent immunoassay kit for detecting phenylethanolamine A includes the following components:

[0088] (1) 96-well opaque white microplate (8 wells x 12 strips) coated with phenylethanolamine A-egg albumin conjugate, vacuum-sealed in an aluminum foil bag;

[0089] (2) 6 bottles of phenylethanolamine A standard solution, the concentrations are respectively:

[0090] 0 ng / mL, 0.04 ng / mL, 0.2 ng / mL, 1.0 ng / mL, 5.0 ng / mL, 25.0 ng / mL;

[0091] (3) phenylethanolamine A-horseradish peroxidase labeled antibody solution;

[0092] (4) Luminescence substrate A solution (luminol and enhancer), luminescence substrate B solution (urea hydrogen peroxide);

[0093] (5) 2 times concentrated sample diluent. Before use, please dilute 1:1 (1 part of concentrated diluent + 1 part of deionized water) to become the working sample diluent, and the diluted working sample diluent is 0.05 mol / L, pH 7.4 glycine-HCl buffer;

[0094] (6) 20 times concentrated washing solut...

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PUM

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Abstract

The invention provides a chemiluminescence immunodetection kit used for detecting phenylethanolamine A, and belongs to the immunological detection field. The kit comprises an opaque white ELISA plate covered with a phenylethanolamine A / carrier protein coupling compound, a phenylethanolamine A standard substance, a phenylethanolamine A / peroxidase labeled antibody working solution, a luminous substrate liquid, a concentrate diluent and a concentrate washing liquid. The phenylethanolamine A / carrier protein coupling compound is obtained by coupling the phenylethanolamine A and a carrier protein through a mixed anhydride method or a carbodiimide method. The concentrate washing liquid comprises 0.05% of tween-20. Compared with a traditional enzyme linked immunosorbent assay method, the kit has characteristics of higher sensitivity, shorter detection time and lower cost and can be used for detection of the residue amount of the phenylethanolamine A in animal urine, blood samples, tissue, viscera, and other samples.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting phenylethanolamine A, which is used for detecting the content or residual amount of phenylethanolamine A in animal-derived foods such as animal tissues, viscera, blood, urine, feed, and feed raw materials. It belongs to the field of immunological detection. Background technique [0002] Phenylethylamine A (Phenylethylamine, PEA), also known as Clonbamine, scientific name is 2-[4-(4-nitrophenyl)butyl-2-ylamino]-1-methoxyphenylethanol, is a A synthetic chemical substance. It can stimulate the growth of animals, strengthen the deposition of protein in animals, improve the level of basal metabolism, and tend to decompose body fat. After people eat meat containing this kind of lean meat extract, they will have nausea, dizziness, limb weakness, hand tremor and other poisoning symptoms, and even induce high blood pressure and heart disease. Long-term consumption may lead to chromosoma...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/68G01N33/577G01N33/543G01N33/535
Inventor 杜道林洪霞
Owner JIANGSU WISE SCI & TECH DEV
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