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Chemiluminescence ELISA detection kit of terbutaline

A technology of chemiluminescent enzyme and terbutaline, which is applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems of complex processing, analysis of many interference factors, long time, etc., and achieve the effect of high sensitivity

Inactive Publication Date: 2009-03-25
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of microbiological methods are: time-consuming and lack of specificity
The disadvantages of thin-layer chromatography are: the operation process is complicated and takes a long time; the operators need to undergo professional training; there are many interference factors affecting the analysis, and the repeatability of the results is poor.
Thin-layer chromatography, radioimmunoassay, high-performance liquid chromatography, color / mass analysis, and liquid / mass analysis have the disadvantages of expensive instruments and equipment, complicated sample pretreatment, time-consuming, laborious, and difficult to popularize , the detection cost is high, especially the radioimmunoassay also needs to be equipped with radioactive sources, which has certain risks

Method used

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  • Chemiluminescence ELISA detection kit of terbutaline
  • Chemiluminescence ELISA detection kit of terbutaline
  • Chemiluminescence ELISA detection kit of terbutaline

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, preparation of immunogen, coated antigen and antibody

[0045] (1) Synthesis of immunogen

[0046] The immunogen was obtained by coupling terbutaline and bovine serum albumin (BSA) by 1,4-butylene ether method. Specifically include the following steps:

[0047] A, Weigh 50 mg of bovine serum albumin (BSA) and dissolve it in 3 mL of 50 mM carbonate (pH10.7) buffer solution, then add 13.8 μL (72.9 μmol) of 1,4-butylene ether (BDE) to the solution at room temperature After 20 hours of reaction, a colorless and clear solution A was obtained;

[0048]B, Weigh 30mg (109.4μmol) terbutaline and dissolve in 1.5mL 50mM carbonate (pH10.7) buffer solution, blow solution A into nitrogen, then add terbutaline solution dropwise into solution A, React at room temperature for 20 hours to obtain a colorless and clear solution;

[0049] C, the reaction solution was transferred to a semi-permeable membrane, and dialyzed with phosphate buffered saline (PBS) (0.01M, pH7.4) ...

Embodiment 2

[0058] Embodiment 2, the establishment of CL-ELISA detection method

[0059] (1) Optimization of antibody and coated antigen concentration (square matrix)

[0060] Serially dilute each coated antigen longitudinally at 40.0 μg / mL, 20.0 μg / mL, 10.0 μg / mL, 5.0 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.625 μg / mL, 0.3125 μg / mL Coat the microtiter plate with 100 μL / well, place overnight at 0-4°C, wash the plate three times with washing solution, and pat dry each time; block with 250 μL / well blocking solution, place at room temperature for 3 hours, wash the plate three times, and pat dry each time ; Add 100 μL / well of a series of diluted antibodies (1:100 to 1:1024000), place at room temperature for 2 hours, wash the plate three times, and pat dry each time; add 100 μL / well of 1:1000 horseradish peroxidase-sheep Anti-rabbit IgG antibody, place at room temperature for 1 hour, wash the plate three times, and pat dry each time; add 100 μL / well of luminescence solution, and measure the luminescen...

Embodiment 3

[0072] Embodiment 3, the chemiluminescent enzyme-linked immunosorbent assay kit that detects terbutaline

[0073] (1) The composition of the chemiluminescent ELISA kit for detecting terbutaline

[0074] A, the solid phase carrier (elisa plate) that is coated with coating antigen (the conjugate of terbutaline and carrier protein);

[0075] B. Terbutaline standard solution: 0.1ng / mL, 0.5ng / mL, 1ng / mL, 5ng / mL, 10ng / mL.

[0076] C. Enzyme-labeled goat anti-rabbit antibody solution: Enzyme-labeled goat anti-rabbit antibody is horseradish peroxidase-goat anti-rabbit IgG stock solution, loaded, and prepared with washing solution to a working concentration of 1:1000 when used.

[0077] D. Terbutaline antibody solution: the polyclonal antibody prepared by immunizing animals with an artificial immune antigen, and dilute the obtained terbutaline antibody with washing solution to a working concentration of 1:600.

[0078] E, luminescent solution: use 0.0001M tris(hydroxymethyl)aminometh...

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Abstract

The invention discloses a chemiluminescence enzyme immunoassay detection reagent kit for terbutaline, which comprises a kit body, an ELIAS plate arranged in the kit body and a reagent in the kit body. The reagent kit is characterized in that each hole of the ELIAS plate is enveloped with an envelope antigen, wherein the envelope antigen is produced by coupling the terbutaline and ovalbumin; and the reagent comprises a terbutaline series standard solution, an enzyme labeled goat anti-rabbit antibody, a terbutaline antibody, a luminescent solution, a washing solution, an envelope solution, and a confining solution. The chemiluminescence enzyme immunoassay detection reagent kit has the characteristics of high sensitivity, simplicity, convenience, quickness, and accuracy; compared with the prior colorimetric ELISA method, the sensitivity can be improved by one order of magnitude; and the reagent kit is expected to play an important role in residue detection of the terbutaline in animal food such as milk, animal tissue, and urine sample.

Description

technical field [0001] The invention relates to an ELISA detection kit, in particular to a chemiluminescent ELISA detection kit for terbutaline. Background technique [0002] Terbutaline belongs to β-agonist, which is a class of chemically synthesized phenylethanolamine derivatives. The mechanism of action of β-stimulants is the same as that of adrenaline and norepinephrine. It can affect the flow and redistribution of nutrients in animals, effectively promote the growth of muscle tissue, reduce carcass fat content, increase lean meat percentage and increase lean meat production. Therefore, it was widely used in Europe and America. Athletes use this drug to increase muscle and lung capacity and shorten the recovery period after high-intensity training. On the other hand, β-agonists have a chemical structure similar to that of adrenaline, and the dosage of such compounds as growth-promoting agents is generally high, generally 5-10 times the therapeutic dosage. Facts have s...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 郗日沫刘中秋丁锴李伟华刘伟尹伟伟
Owner SHANDONG UNIV
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