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Thyroglobulin chemical luminescence detection kit

A technology for detection of thyroglobulin and chemiluminescence, which is applied in chemiluminescence/bioluminescence, biological testing, and analysis through chemical reactions of materials, etc., which can solve the problems of low sensitivity and high cost of reagents

Active Publication Date: 2018-08-24
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In China, the detection of thyroglobulin is mainly based on chemiluminescence immunoassay in clinical practice. However, due to the lack of domestic thyroid detection kits with independent intellectual property rights, low sensitivity and high cost of reagents, they are generally only available in China. It has been carried out in tertiary hospitals, but less in hospitals below the tertiary level. Therefore, it is necessary to develop a detection technology with high sensitivity, good accuracy and low detection cost for thyroid detection.

Method used

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  • Thyroglobulin chemical luminescence detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1: Preparation of avidinylated magnetic particles (reagent 1) coated with anti-thyroglobulin antibody 1. Preparation of biotinylated anti-thyroglobulin antibody

[0020] Take 1.0 mg of anti-thyroglobulin antibody, dilute it with phosphate buffer to 1.0 mg / mL, then add 3.33 μL of biotin ester with a concentration of 10 mM / L, react at room temperature for 30 min, and then add 10 μL of Tris solution with a concentration of 1 M, The reaction was carried out at room temperature for 10 min, and finally dialyzed with 20 mM phosphate buffer with a pH of 7.4 for 16-24 hours, and the biotinylated anti-thyroglobulin antibody was obtained by taking the dialyzed liquid.

[0021] 2. Biotinylated Antibody Coated Avidin Magnetic Microparticles

[0022] Take 100 mg of avidinylated magnetic particle suspension, magnetically separate the supernatant, and use ligation buffer (20 mM PBS, 10 g / L BSA, 20 g / L mannitol, 10 mL / L glycerol, 0.1 mL / L TritonX- 100, 3mL / L proclin-300, 20mg / L...

Embodiment 2

[0024] Example 2: Preparation of acridine ester-labeled anti-thyroglobulin antibody (reagent 2)

[0025] Take 0.5 mg of anti-thyroglobulin antibody, dilute it to 1.0 mg / mL with 20 mM phosphate buffer, then add 2.5 μL of acridine ester with a concentration of 10 mM / L, mix at room temperature for 30 min in the dark, and then add 50 μL of 10 mg concentration / mL of lysine solution, mixed for 10 min in the dark at room temperature, and then purified by centrifugation with a desalting column. Collect the liquid in the centrifuge tube to obtain the acridine ester-labeled anti-thyroglobulin antibody. The acridine ester-labeled anti-thyroglobulin antibody was prepared with marker buffer (20mM PBS, 10g / L casein, 10g / L trehalose, 10mL / L glycerol, 0.1mL / L TritonX-100, 3mL / L proclin-300, 20mg / L 4-aminoantipyrine) was diluted at a ratio of 1 / 1000 to prepare the working solution of acridinium ester marker.

[0026] Following this method, Reagent 2 comprising 2.0 μg / mL, 4.0 μg / mL and 6.0 μ...

Embodiment 3

[0027] Example 3: Preparation of Thyroglobulin Calibrator

[0028] Diluent with calibrator (20mM PBS, 10g / L casein, 20g / L trehalose, 10mL / L glycerol, 0.1mL / L TritonX-100, 3mL / L proclin-300, 20mg / L 4-aminoantibiotic Billing) thyroglobulin antigen was formulated into Calibrator 1 and Calibrator 2 at concentrations of 1 ng / mL, 100 ng / mL. Thyroglobulin calibrators are traceable to Certified Reference Materials (CRM) 457 of the European Community Bureau for Reference Materials (BCR).

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Abstract

The invention discloses a thyroglobulin chemical luminescence detection kit. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 is prepared from buffer liquor, magnetic particles, biotin-labelled anti-thyroglobulin monoclonal antibody and polyhydroxy-alcohol; and the reagent 2 is prepared from buffer liquor, an acridinium ester labeled anti-thyroglobulin monoclonal antibody and polyhydroxyl sugar. The thyroglobulin chemical luminescence detection kit is high in sensitivity, is good in accuracy, and can reduce detection cost.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a thyroglobulin chemiluminescence detection kit. Background technique [0002] Thyroglobulin (TG) is an iodinated glycoprotein secreted by the thyroid follicular epithelium with a molecular weight of about 660KD. Thyroglobulin is synthesized by thyroid cells under the action of thyroid stimulating hormone (TSH) and transported into thyroid follicles, where it is stored as a prerequisite for thyroxine and iodothyronine. Thyroglobulin is hydrolyzed by thyroid peroxidase (TPO) to generate thyroxine (T4) and triiodothyronine (T3) which are released into the blood. TG levels in serum can be affected by a variety of thyroid-related diseases, including Hashimoto's disease (thyroiditis), Graves' disease (toxic diffuse goiter), subacute thyroiditis, hyperthyroidism, and thyroid tumors. In patients with congenital thyroid dysfunction, detection of thyroglobulin levels can iden...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/68G01N33/577
CPCG01N21/76G01N33/577G01N33/6893G01N2800/046
Inventor 吴吉兵吴婷徐雨田君喜龙腾镶
Owner SICHUAN MACCURA BIOTECH CO LTD
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