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Tetracycline chemiluminescence detecting kit and preparation method thereof

A chemiluminescence detection and chemiluminescence technology, which is applied in chemiluminescence/bioluminescence, analysis through chemical reactions of materials, and measurement devices, can solve problems such as large error in results, low sensitivity, and poor specificity of microbial methods, and achieve The photon yield does not decrease, the reagent stability is good, and the effect of less interference factors

Inactive Publication Date: 2017-10-24
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 microbiological method has poor specificity, cumbersome operation, long detection cycle and large error in results
CN 1707265 A (December 2005) discloses a test kit for detecting tetracycline by enzyme-linked immunoassay. The disadvantage of using enzyme-linked immunoassay for detection is that horseradish peroxidase or alkaline phosphate markers are easily inactivated and color-developed The substrate is easy to decompose when exposed to light, has low sensitivity, and has a certain cross-reaction to compounds with similar structures, resulting in inaccurate test results, and is only suitable for the detection and identification of trace substances
CN 102401818 A (April 2012) discloses a method for detecting tetracycline by high-performance liquid chromatography. The disadvantage of using high-performance liquid chromatography is that the sample pretreatment is complicated, laborious and time-consuming, and the detection cost is high
CN 103105490 A (November 2011) discloses a kit for colloidal gold detection of tetracycline. The disadvantage of using colloidal gold method for detection is that the final detection result is characterized by a color visible to the naked eye, with large errors and cumbersome operations. More processes, more prone to errors, low sensitivity

Method used

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preparation example Construction

[0023] The present invention provides a tetracycline chemiluminescence detection kit and preparation method, wherein, the chemiluminescence detection kit provided by the present invention can adopt tetracycline antibody-coupled magnetic particles, acridinium ester-labeled tetracycline, or can adopt Tetracycline antigen-coupled magnetic particles, acridinium ester-labeled tetracycline antibody. The kit also includes tetracycline 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 process of preparing the magnetic p...

Embodiment 1

[0031] Example 1: The composition and preparation method of a tetracycline chemiluminescence detection kit 1

[0032] 1. Assembly of Kit 1

[0033] acridinium ester-labeled tetracycline antigen;

[0034] Tetracycline antibody coupled to magnetic particles;

[0035] Tetracycline series calibrator solutions, the concentrations are: 0 μg / L, 0.02 μg / L, 0.1 μg / L, 0.5 μg / L, 2.5 μg / L, 12.5 μg / L, 62.5 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 Suspension Conjugated to Tetracycline 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 f...

Embodiment 2

[0056] Example 2: The composition and preparation method of a tetracycline chemiluminescence detection kit 2

[0057] 1. Assembly of Kit 2

[0058] Acridinium ester-labeled tetracycline antibody;

[0059] Tetracycline antigen coupled to magnetic particles;

[0060] Tetracycline series calibrator solutions, the concentrations are: 0 μg / L, 0.02 μg / L, 0.1 μg / L, 0.5 μg / L, 2.5 μg / L, 12.5 μg / L, 62.5 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 particle suspension coupled with tetracycline 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 5 m...

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Abstract

The invention discloses a tetracycline chemiluminescence detecting kit and a preparation method thereof. The kit comprises an acridinium ester marked tetracycline antigen or antibody, a magnetic particle coupled antigen or antibody, a tetracycline series standard substance solution, a chemiluminescence pre-excited solution A, a chemiluminescence pre-excited solution B and a cleanout fluid. The kit integrates a chemiluminescence technology and immune magnetic particles, and a reaction system approximate to a homogeneous phase is provided. Compared with the prior art, the kit 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 tetracycline immunochemiluminescent detection kit and a preparation method thereof. Background technique [0002] Tetracycline is a derivative of tetracene or naphthocene, which is a kind of spectrum antibacterial drug. It is an antibiotic extracted from the culture solution of Streptomyces or semi-synthesized. It is named for its molecule containing 4 rings. Adding tetracycline to aquatic products, milk, eggs, royal jelly, etc. can be used to inhibit infections caused by various bacteria and rickettsia, chlamydia, mycoplasma, etc. However, tetracycline drugs are mainly filtered out through the glomerulus. These drugs contain many hydroxyl groups, enol hydroxyl groups and carbonyl groups, and can form insoluble chelates with various metal ions under neutral conditions. Such as forming insoluble calcium or magnesium salts with calcium or magnesium ions, and forming red com...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/76
CPCG01N33/54326G01N21/76
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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