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Chemiluminescence detection kit

A technology of chemiluminescence detection and kits, which is applied in chemiluminescence/bioluminescence, analysis by making materials undergo chemical reactions, and measurement devices, which can solve the problems of high reagent cost and low sensitivity

Active Publication Date: 2018-09-07
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The limitations of the washing conditions described in the current chemiluminescence immunoassay kits must be equipped with expensive equipment for the experiment, and the experimental operation steps of the current kits are cumbersome. Some chemiluminescence detection kits have low sensitivity and high reagent costs. Currently, they are generally only in the top three It has been carried out in hospitals, but less in hospitals below the top three. Therefore, it is necessary to develop a chemiluminescence kit with high sensitivity, good accuracy and low detection cost.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0017] Embodiment one: the preparation and sensitivity experiment of renin chemiluminescence detection kit

[0018] 1. Preparation of avidin-coated magnetic particles (reagent 1) coated with renin monoclonal antibody

[0019] 1. Preparation of Biotinylated Renin Monoclonal Antibody

[0020] Take 1.0 mg of renin monoclonal antibody, dilute to 0.5 mg / mL with phosphate buffer, then add 13.3 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 min, and finally dialyze with 20 mM phosphate buffer solution with pH 7.4 for 16-24 hours, and obtain the biotinylated renin monoclonal antibody by taking the dialyzed liquid.

[0021]2. Biotinylated antibody coated avidin magnetic particles

[0022] Take 50 mg of avidinized magnetic particle suspension, magnetically separate to remove the supernatant, and use ligation buffer (20mM PBS, 10g / LBSA, 20g / L man...

Embodiment 2

[0051] Embodiment two: preparation and sensitivity experiment of thyroglobulin chemiluminescence detection kit

[0052] 1. Preparation of avidin-coated magnetic particles (reagent 1) coated with anti-thyroglobulin antibody

[0053] 1. Biotinylated anti-thyroglobulin antibody preparation

[0054] Take 1.0mg of anti-thyroglobulin antibody, dilute it to 1.0mg / mL with phosphate buffer, then add 3.33μL of biotin ester with a concentration of 10mM / L, react at room temperature for 30min, then add 10μL of Tris solution with a concentration of 1M, React at room temperature for 10 minutes, and finally dialyze with 20 mM phosphate buffer solution with pH 7.4 for 16-24 hours, and obtain the biotinylated anti-thyroglobulin antibody by taking the dialyzed liquid.

[0055] 2. Biotinylated antibody coated avidin magnetic particles

[0056] Take 100 mg of avidinized magnetic particle suspension, magnetically separate to remove the supernatant, and use ligation buffer (20mM PBS, 10g / L BSA, 20...

Embodiment 3

[0086] Example Three: Preparation and Sensitivity Experiment of Hepatitis B Pre-S1 Antigen Chemiluminescence Detection Kit

[0087] 1. Preparation of avidin-coated magnetic particles (reagent 1) coated with anti-PreS1-Ag monoclonal antibody

[0088] 1. Preparation of biotinylated anti-PreS1-Ag monoclonal antibody

[0089] Take 1.0mg anti-PreS1-Ag monoclonal antibody, dilute it to 1.0mg / mL with phosphate buffer, then add 13.3μL biotin ester with a concentration of 10mM / L, react at room temperature for 30min, and then add 10μL Tris with a concentration of 1M The solution was mixed at room temperature for 10 minutes to terminate the reaction, and finally dialyzed with 20 mM phosphate buffer solution with pH 7.4 for 16-24 hours, and the dialyzed liquid was taken to obtain biotinylated anti-PreS1-Ag monoclonal antibody.

[0090] 2. Biotinylated antibody coated avidin magnetic particles

[0091] Take 50mg of avidinized magnetic particle suspension, magnetically separate to remove ...

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Abstract

The invention discloses a chemiluminescence detection kit. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 is prepared from a buffer solution, magnetic particles, a first antibodyof a biotin-labeled to-be-detected antigen, and polyhydric alcohol; and the reagent 2 is prepared from a buffer solution, a second antibody of an acridinium ester-labeled to-be-detected antigen, andpolyhydroxy saccharide. The chemiluminescence detection kit is high in sensitivity and good in accuracy, and can lower the detection cost.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a chemiluminescence detection kit. Background technique [0002] The chemiluminescence detection kit utilizes two detection substances (antibodies, antigens or other molecules with affinity for specific analytes) to detect a certain substance in the sample. One of the detection substances is coated on the solid surface (hereinafter referred to as solid phase), and the other detection substance is labeled with enzyme or chemiluminescent substance (hereinafter referred to as label). The current analytical method for the use of chemiluminescent immunoassay kits is to use a luminescent agent as a marker of an antibody or antigen, and directly detect the content of the antigen or antibody in the specimen to be tested through a luminescence reaction. The principle steps of the kit are as follows: ①Add the sample to be tested into the reaction cup, keep it warm for a period ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/543
CPCG01N21/76G01N33/54326
Inventor 徐雨黄成毅田君喜龙腾镶
Owner SICHUAN MACCURA BIOTECH CO LTD
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