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Adrenocorticotropic hormone (ACTH) chemiluminescence detection kit

A chemiluminescence detection, adrenal cortex technology, used in chemiluminescence/bioluminescence, biological testing, analysis by chemical reaction of materials, etc., can solve the problems of high reagent cost and low sensitivity

Active Publication Date: 2018-09-18
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In China, the detection of adrenocorticotropic hormone is mainly based on chemiluminescent immunoassay in clinical practice. However, due to the fact that domestic adrenocorticotropic hormone detection kits with independent intellectual property rights are still relatively small, and the sensitivity is low and the cost of reagents is high. At present, it is generally only carried out in tertiary hospitals, and less in hospitals below the tertiary level. Therefore, it is necessary to develop a detection technology for corticotropic hormone with high sensitivity, good accuracy and low detection cost.

Method used

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  • Adrenocorticotropic hormone (ACTH) chemiluminescence detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of avidinized magnetic particles (reagent 1) coated with corticotropin monoclonal antibody

[0019] 1. Preparation of biotinylated corticotropin monoclonal antibody

[0020]Take 1.0 mg of corticotropin monoclonal antibody, dilute to 0.5 mg / mL with phosphate buffer (20 mM pH7.4), then add 25 μL of biotin ester with a concentration of 2.5 mM / L, react at room temperature for 30 min, and then Add 20 μL of Tris solution with a concentration of 1M, react at room temperature for 10 minutes, and finally dialyze with phosphate buffer (20Mm pH 7.4) for 16-24 hours, and obtain the biotinylated corticotropin monoclonal antibody by taking the dialyzed liquid.

[0021] 2. Biotinylated antibody coated avidin magnetic particles

[0022] Take 100 mg of avidinized magnetic particle suspension, magnetically separate to remove the supernatant, and use ligation buffer (20mM PBS, 10g / L BSA, 20g / L mannitol, 10mL / L glycerol, 0.1mL / L TritonX- 100, 1mL / L proclin-300, 20m...

Embodiment 2

[0023] Example two: Preparation of acridinium ester-labeled corticotropin monoclonal antibody (reagent 2)

[0024] Take 1.0mg of corticotropin monoclonal antibody, dilute to 1.0mg / mL with phosphate buffer (20mM pH7.4), then add 100μL of acridinium ester with a concentration of 2mM / L, and mix and react at room temperature for 30min in the dark , and then add 100 μL of lysine solution with a concentration of 10 mg / mL, mix and react for 10 min at room temperature in the dark, and then desalt and purify by centrifugation with a desalting column. Collect the liquid in the centrifuge tube to obtain the acridinium ester-labeled corticotropin monoclonal antibody. Then the acridinium ester-labeled corticotropin monoclonal antibody obtained above was diluted to a concentration of 2 μg / mL with a label diluent to obtain reagent 2. The marker diluent is 20mM pH6.0 2-morphine ethanesulfonic acid solution, containing 9g / L sodium chloride, 10g / L casein, 10g / L trehalose, 10mL / L glycerol, 0.1m...

Embodiment 3

[0025] Embodiment three: preparation of adrenocorticotropic hormone calibrator

[0026] The corticotropin antigen was prepared with the calibrator diluent to a concentration of 0pg / mL, 5pg / mL, 25pg / mL, 50pg / mL, 125pg / mL, 500pg / mL, 1000pg / mL, 2000pg / mL. Calibrator diluent is 20mM pH6.0 2-morphine ethanesulfonic acid solution, containing 9g / L sodium chloride, 10g / L casein, 20g / L trehalose, 10mL / L glycerol, 0.1mL / L TritonX- 100, proclin-300 1mL / L and 20mg / L 4-aminoantipyrine.

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Abstract

The invention discloses an adrenocorticotropic hormone (ACTH) chemiluminescence detection kit, which comprises a reagent (1) and a reagent (2). The reagent (1) comprises a buffer solution, magnetic particles, a biotin labeled anti-ACTH monoclonal antibody, and polyhydric alcohol. The reagent (2) comprises a buffer solution, an acridinium ester labeled anti-ACTH monoclonal antibody and polyhydroxylsaccharides. The provided kit has the advantages of good stability, high sensitivity, good accuracy, simple and convenient operation, and low cost, and is not influenced by rheumatoid factors.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a corticotropin chemiluminescence detection kit. Background technique [0002] Adrenocorticotropic hormone (ACTH) is a peptide hormone composed of 39 amino acids with a molecular weight of about 4.5kD. Adrenocorticotropic hormone is produced in the pituitary gland and acts on the adrenal cortex to promote the proliferation of adrenal cortical tissue and the production and secretion of corticosteroids. The secretion of corticotropin is regulated by many factors such as hypothalamic corticotropin releasing factor (CRH), cortisol, and self-negative feedback. [0003] Corticotropin testing is valuable in the differential diagnosis of adrenal insufficiency and hypersecretion. Increased corticotropin levels seen in: Addison disease (primary adrenal insufficiency), corticotropin insensitivity syndrome and corticotropin receptor abnormalities, Cushing disease secondary to pi...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N21/76
CPCG01N21/76G01N33/74G01N2333/695
Inventor 黄燕玲赵浩瀚郭鹏田君喜龙腾镶
Owner SICHUAN MACCURA BIOTECH CO LTD
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