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Chemiluminescence detection kit for doxycycline, and preparation method of chemiluminescence detection kit

A chemiluminescence detection, doxycycline technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve problems such as large error in results, large influence of enzymatic environment, and low sensitivity , to achieve the effect of no reduction in photon yield, good reagent stability and low background luminescence

Inactive Publication Date: 2017-12-29
太原瑞盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specificity of the microbiological method is poor, the detection period is long and the result error is large
ELISA is not sensitive enough and is only suitable for the detection and identification of trace substances
High performance liquid chromatography sample pretreatment is complex, laborious and time-consuming, and the detection cost is high
CN 102213723 A (2010) discloses a kit for detecting doxycycline by the colloidal gold method. The disadvantage of using the colloidal gold method for detection is low sensitivity
CN 106556702 A (November 2016) disclosed a horseradish peroxidase-labeled doxycycline antigen detection kit for doxycycline. The disadvantage of using horseradish peroxidase for detection is that the enzymatic process is greatly affected by the environment. : For example, disinfectant, pH, and ions will all affect it, thereby affecting the measurement results
In addition, enzymatic chemiluminescence has the following disadvantages: such as long time for the substrate to reach the plateau, low sensitivity, etc.

Method used

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Examples

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preparation example Construction

[0023] The present invention provides a chemiluminescent detection kit for doxycycline and a preparation method thereof, wherein, the chemiluminescent detection kit for doxycycline provided by the present invention can use doxycycline antibody-coupled magnetic particles, acridinium ester To label doxycycline, you can also take doxycycline antigen-coupled magnetic particles, and acridinium ester-labeled doxycycline antibody. The kit also includes doxycycline calibrator, chemiluminescence pre-excitation solution A, chemiluminescence excitation solution B and cleaning solution.

[0024] Specifically, in the process of preparing magnetic particle coupling in the present invention, the buffer solution for coupling antigen is a buffer solution with a pH value of 5.0 and a concentration of 0.1 mol / L MES; the buffer solution for coupling antibody is a buffer solution with a pH value of 5.0 6.0. Buffer solution with a concentration of 0.1 mol / L MES.

[0025] Specifically, in the proce...

Embodiment 1

[0031] Example 1: The composition and preparation method of a chemiluminescent detection kit 1 for doxycycline

[0032] 1. Assembly of Kit 1

[0033] Doxycycline antigen labeled with acridinium ester;

[0034] Doxycycline antibody coupled to magnetic particles;

[0035] Doxycycline series calibrator solution, the concentrations are: 0 μg / L, 0.1 μg / L, 0.3 μg / L, 0.9 μg / L, 2.7 μg / L, 8.1 μg / L, 24.0 μg / L;

[0036] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0037] Cleaning solution, including the following components: Tris-HCl with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl and 0.05% Tween-20 with a concentration of 0.15 mmol / L.

[0038] 2. Preparation of Magnetic Microparticle Suspensions Conjugated to Doxycycline Antibody

[0039](1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add a certain amount of 0.1 mol / L MES buffer, vortex and mix, place on a magnetic stand, and let it stand ...

Embodiment 2

[0056] Example 2: The composition and preparation method of a chemiluminescent detection kit 2 for doxycycline

[0057] 1. Assembly of Kit 2

[0058] Acridinium ester-labeled doxycycline antibody;

[0059] Doxycycline antigen coupled to magnetic particles;

[0060] Doxycycline series calibrator solution, the concentrations are: 0 μg / L, 0.1 μg / L, 0.3 μg / L, 0.9 μg / L, 2.7 μg / L, 8.1 μg / L, 24.0 μg / L;

[0061] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0062] Cleaning solution, including the following components: Tris-HCl with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl and 0.05% Tween-20 with a concentration of 0.15 mmol / L.

[0063] 2. Preparation of Magnetic Microparticle Suspension Conjugated with Doxycycline Antigen

[0064] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add a certain amount of 0.1 mol / L MES buffer, vortex and mix, place on a magnetic stand, and let it stand for ...

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Abstract

The invention discloses a chemiluminescent detection kit for doxycycline and a preparation method thereof. The kit includes: acridinium ester-labeled doxycycline antigen or antibody, magnetic particle-coupled antibody or antigen, doxycycline series standard solution, chemiluminescence pre-excitation solution A, chemiluminescence excitation solution B, and cleaning solution. The kit of the invention combines the chemiluminescent technology with the immunomagnetic particle, and provides a nearly homogeneous reaction system. Compared with the prior art, the kit of the invention has the advantages of high sensitivity, strong specificity, short reaction time and the like.

Description

technical field [0001] The invention belongs to the field of food safety detection, in particular to a doxycycline immunochemiluminescence detection kit and a preparation method thereof. Background technique [0002] Doxycycline (DC) is a tetracycline drug whose main component is doxycycline hydrochloride. It is generally used as an additive in dairy products, honey, organic fertilizers, etc. to prevent and treat diseases caused by parasites, mycoplasma, bacteria, etc., and can also play a very good antibacterial and antibacterial effect. In the long-term treatment practice, people found that doxycycline is a good antibacterial drug, but if this type of antibiotic is often abused, the parasite will acquire stronger drug resistance than before. In addition, after these antibiotics are enriched in animals, they will enter the human body through the food chain and endanger human health. The products exported by our country have been boycotted by the European Union, the United...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/76
CPCG01N33/9446G01N21/76G01N33/54326
Inventor 曹晶刘丽青胡雪婷常燕杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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