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Chemical luminescent detection kit of thyroglobulin and preparation method thereof

A technology for thyroglobulin and chemiluminescence detection, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, etc. Fast and convenient, improved sensitivity, high accuracy

Inactive Publication Date: 2018-03-23
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

Among them, CN 106932573A (2017.07) discloses an immune colloidal gold test strip for detecting thyroglobulin and its preparation method. The method uses colloidal gold to label thyroglobulin antibody. The detection of the colloidal gold method is characterized by the color visible to the naked eye. Large error, cumbersome operation, many processes, low sensitivity, low precision and other disadvantages; CN 106706927 A (2017.05) discloses a thyroglobulin ELISA diagnostic kit, which adopts ELISA method for detection, enzyme The antigen and antibody of the immunoassay method are combined on the surface of the solid phase (ELISA plate reaction well), which has the disadvantages of low sensitivity, difficulty in realizing full automation, and long detection time; CN 105510593 A (2016.04) discloses a thyroid bulb Protein (TG) test kit and its detection method. The kit uses alkaline phosphatase as a marker. The disadvantage of using alkaline phosphatase for labeling is that the enzymatic process is greatly affected by the environment: such as disinfectant, pH, and ions. It has an impact and weak stability, thus affecting the measurement results

Method used

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preparation example Construction

[0024] The present invention provides a thyroglobulin chemiluminescence reagent kit and a preparation method thereof. In order to make the purpose, technical scheme and effect of the present invention clearer and clearer, the present invention will be further described in detail below.

[0025] The invention provides a thyroglobulin chemiluminescence reagent kit, which uses magnetic separation chemiluminescence technology as a detection means and combines acridinium ester labeling technology for detection. The kit includes: magnetic particle-coupled thyroglobulin capture antibody, acridinium ester-labeled thyroglobulin detection antibody, thyroglobulin calibrator solution, chemiluminescence pre-excitation solution A, chemiluminescence excitation solution B, and cleaning solution.

[0026] Specifically, the magnetic particles of the present invention can be directly coupled with the thyroglobulin capture antibody, or the magnetic particles can be coupled with streptavidin, and t...

Embodiment 1

[0035] Embodiment 1: The formation and preparation method of a kind of detection thyroglobulin chemiluminescence kit, comprises the following steps:

[0036] 1. Kit set-up

[0037] Construct a chemiluminescence detection kit for thyroglobulin so that it contains the following components:

[0038] Thyroglobulin monoclonal antibody coupled to carboxyl magnetic beads;

[0039] Acridinium ester-labeled thyroglobulin monoclonal antibody;

[0040] Thyroglobulin series calibrator solution;

[0041] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0042] Washing fluid.

[0043] 2. Preparation of Magnetic Microparticle Suspension Conjugated with Thyroglobulin Monoclonal Antibody

[0044] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add 200 μL of 0.1 mol / L MES (pH 6.0) buffer, vortex and mix, place on a magnetic stand, and let the magnetic particles stand for 5 min to make the magnetic particles Separate from t...

Embodiment 2

[0062] Example 2: Detection of Thyroglobulin in Actual Samples

[0063] The operating procedure of the thyroglobulin quantitative detection kit of the present invention is as follows:

[0064] Detection of the kit

[0065] (1) Add 50 μL of the sample to be tested into the cuvette, then add 150 μL of magnetic particle coupling suspension, shake and mix, and incubate at 37°C for 8 min.

[0066] (2) Separation, washing 3 times, fully shaking the washed reaction vessel to disperse the magnetic particles.

[0067] (3) Add 150 μL of acridinium ester marker into the cuvette, shake to mix, and incubate at 37°C for 7 min.

[0068] (4) Separation, washing 3 times, fully shaking the washed reaction vessel to disperse the magnetic particles.

[0069] (5) Add 100 μL of chemiluminescence pre-excitation solution A, add 100 μL of chemiluminescence excitation solution B after 1 s, and measure its relative luminous intensity. The content of thyroglobulin in the sample is proportional to its ...

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Abstract

The invention discloses a thyroglobulin chemiluminescence detection kit and a preparation method thereof. The kit includes: magnetic particles coupled with thyroglobulin capture antibody, acridine ester-labeled thyroglobulin detection antibody, thyroglobulin calibrator solution, cleaning solution, chemiluminescence pre-excitation solution A, and chemiluminescence excitation solution B . The kit of the invention uses the magnetic separation chemiluminescence technology as the detection method, and combines the acridinium ester labeling technology at the same time. Compared with the prior art, the kit has the advantages of high sensitivity, fast and convenient detection, high accuracy, good repeatability, strong specificity and the like. .

Description

technical field [0001] The invention belongs to the technical field of immune detection and analysis, and in particular relates to a thyroglobulin chemiluminescent detection kit and a preparation method thereof. Background technique [0002] Thyroglobulin (TG) is a large glycoprotein (MW 660,000) present in thyroid follicular cells. Each TG contains about 2 thyroxine (T4) and 0.5 triiodothyronine (T3). These two hormones are produced by the iodination and coupling of some tyrosine in the TG molecule. The main physiological role of TG is to serve as a substrate for the synthesis of thyroid hormones, store iodide ions, and transport and release thyroid hormones. TG has multiple effects on thyroid cells. On the one hand, it can promote the proliferation of thyroid follicular epithelial cells at low concentrations, and can inhibit the growth of thyroid cells at high concentrations; on the other hand, TG can also antagonize the proliferation of thyroid cells at high concentratio...

Claims

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Application Information

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IPC IPC(8): G01N21/76
CPCG01N21/76
Inventor 刘丽青曹晶常燕胡雪婷杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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