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Hepatitis B PreS1 antigen chemiluminescence detection kit

A chemiluminescence detection and kit technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problems of low sensitivity, high reagent cost, large error, etc.

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

The plate chemiluminescence method is similar to the enzyme-linked immunosorbent method, with a low degree of automation and mostly manual operations. The detection results have large errors due to human factors and low sensitivity; the magnetic particle chemiluminescence method uses fully automatic instruments for detection, and the degree of automation Higher and more stable results, but the kits currently used in clinical practice generally have problems of low sensitivity and insufficient low-end resolution, and multi-point calibrator is often used for calibration, which increases the cost of reagent testing and the burden on patients
[0006] In China, pre-S1 antigen detection is mainly based on chemiluminescent immunoassay in clinical practice. However, due to the fact that there are still relatively few domestic pre-S1 antigen detection kits with independent intellectual property rights, and the sensitivity is low and the cost of reagents is high. It is only carried out in tertiary hospitals, and less in hospitals below the tertiary level. Therefore, a detection technology with high sensitivity, good specificity and low detection cost for pre-S1 antigen detection needs to be developed

Method used

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  • Hepatitis B PreS1 antigen chemiluminescence detection kit
  • Hepatitis B PreS1 antigen chemiluminescence detection kit
  • Hepatitis B PreS1 antigen chemiluminescence detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1: Preparation of avidinized magnetic particles (reagent 1) coated with anti-PreS1-Ag monoclonal antibody 1. Preparation of biotinylated anti-PreS1-Ag monoclonal antibody

[0020] 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.

[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 (20 mM PBS, 10 g / LBSA, 20 g / L mannitol, 10 mL / L glycerol, 20 mg / L 4-aminoaminophen Tipyri...

Embodiment 2

[0024] Embodiment two: the anti-HBsAg polyclonal antibody (reagent 2) preparation of acridinium ester labeling

[0025] Take 1.0mg of anti-HBsAg polyclonal antibody, dilute it to 1.0mg / mL with 20mmol / L phosphate buffer, then add 100μL of acridinium ester with a concentration of 2mmol / L, mix and react at room temperature for 30min in the dark, then add 100μL of 10mg / mL lysine solution, mix and react in the dark at room temperature for 10min, and then desalt and purify by centrifugation with a desalting column. Collect the liquid in the centrifuge tube to obtain the acridinium ester-labeled anti-HBsAg polyclonal antibody and add an equal volume of glycerol to store it below -20°C.

[0026] Take 100mL of buffer solution (20mM phosphate, 9.00g / L sodium chloride, 10.00g / L casein, 10.00g / L trehalose, 20mg / L 4-aminoantipyrine, 0.1mL / L Triton X- 100, 10mL / L glycerol, 1.00mL / L proclin-300) was added thereto with 10 μL of the above-mentioned acridinium ester-labeled anti-HBsAg polyclon...

Embodiment 3

[0027] Embodiment three: Preparation of PreS1-Ag calibrator

[0028] Hepatitis B virus pre-S1 antigen buffer (20mM pH7.4PBS, 10g / L casein, 20g / L trehalose, 10mL / L glycerol, 0.5mL / L Tween, 1mL / L proclin-300) Prepared to a concentration of 0 μg / mL and 10 μg / mL, namely calibrator 1 and calibrator 2.

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Abstract

The invention discloses a hepatitis B PreS1 antigen chemiluminescence detection kit. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 is prepared from buffer liquid, magnetic particles, biotin marked anti-hepatitis B PreS1 antigen monoclonal antibody and polyhydroxy alcohol; the reagent 2 is prepared from buffer liquid, acridinium ester marked anti-hepatitis B PreS1 antigen monoclonal antibody and polyhydroxy saccharide. The kit has the advantages that in the use process, the signal background is low; the signal intensity is obviously improved; the sensitivity, the accuracyand the repeatability of the kit are obviously improved.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a chemiluminescence detection kit for hepatitis B pre-S1 antigen. Background technique [0002] Hepatitis B is an infectious disease that seriously endangers human health. About 2 billion people in the world have been infected with hepatitis B virus (HBV), of which 350 million people are chronically infected with HBV. [0003] There are four gene regions encoding HBV protein: S gene region, C gene region, P gene region and X gene region. The S gene region is composed of S gene, pre-S2 gene and pre-S1 gene, which encode HBsAg, pre-S2 protein, pre-S1 protein and polymeric human serum albumin receptor respectively. HBV pre-S1 antigen (PreS1-Ag) mainly exists on the surface of intact Dane granules and cast granules, is one of the surface antigen protein components encoded by the pre-S1 gene, and is generally co-expressed with HBsAg in the serum of HBV-infected patients. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/577G01N33/576
CPCG01N21/76G01N33/5764G01N33/577
Inventor 樊琦昊赵燕莉徐雨田君喜龙腾镶
Owner SICHUAN MACCURA BIOTECH CO LTD
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