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Hepatitis B surface antigen chemiluminiscence immune detection reagent kit and application thereof

A technology of chemiluminescence immunization and HBsAg, applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, biological testing, etc., can solve the problem of low detection ability of HBsAg mutant strains, and improve Sensitivity and accuracy, high sensitivity, high affinity effects

Active Publication Date: 2018-07-31
SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is: in order to solve the technical problem that the existing kits have low detection ability to the mutant strain of hepatitis B surface antigen, a kind of hepatitis B surface antigen chemiluminescent immunoassay kit and its application are provided

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  • Hepatitis B surface antigen chemiluminiscence immune detection reagent kit and application thereof
  • Hepatitis B surface antigen chemiluminiscence immune detection reagent kit and application thereof
  • Hepatitis B surface antigen chemiluminiscence immune detection reagent kit and application thereof

Examples

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Embodiment 1

[0025] This embodiment provides a hepatitis B surface antigen chemiluminescent immunoassay kit, comprising: a magnetic particle reagent indirectly connected to anti-HBsAg antibody 1, a solution of HBsAg antibody 2 labeled with a chemiluminescent marker, a chemiluminescent substrate solution, and a cleaning solution;

[0026] The anti-HBsAg antibody 1 is composed of an anti-S protein monoclonal antibody 1 and a polyclonal antibody Fab fragment with a molar ratio of 1:0.2;

[0027] The anti-HBsAg antibody 2 is composed of an anti-S protein monoclonal antibody 2 and a polyclonal antibody Fab fragment with a molar ratio of 1:2;

[0028] The polyclonal antibody Fab fragment is prepared and purified by the following method: the polyclonal antibody and the specific Fab processing enzyme are reacted at 37°C for 24 hours in a 0.2M Tris-HCl buffer system with a pH of 7.2 at a volume ratio of 1:0.5, Obtain the digested product; pass the digested product through a protein G affinity colum...

Embodiment 2

[0033] This embodiment provides a hepatitis B surface antigen chemiluminescent immunoassay kit, which is basically the same as the kit in Example 1, except that:

[0034] The anti-HBsAg antibody 1 is composed of an anti-S protein monoclonal antibody 1 and a polyclonal antibody Fab fragment with a molar ratio of 1:1;

[0035] The anti-HBsAg antibody 2 is composed of an anti-S protein monoclonal antibody 2 and a polyclonal antibody Fab fragment with a molar ratio of 1:0.5;

[0036] The polyclonal antibody Fab fragment is prepared and purified by the following method: the polyclonal antibody and the specific Fab processing enzyme are reacted in a 0.2M Tris-HCl buffer system with a pH of 7.2 at a volume ratio of 1:1 at 37°C for 12 hours, Obtain the digested product; pass the digested product through a protein G affinity column to collect the effluent protein P1, fully equilibrate the protein G affinity column with 0.2M PBS buffer at pH 7.2, then use 0.1M HCl- Glycine was eluted t...

Embodiment 3

[0040] This embodiment provides a hepatitis B surface antigen chemiluminescent immunoassay kit, which is basically the same as the kit in Example 1, except that:

[0041] The anti-HBsAg antibody 1 is composed of an anti-S protein monoclonal antibody 1, an anti-pre-S2 protein monoclonal antibody and a polyclonal antibody Fab fragment with a molar ratio of 1:0.2:1;

[0042] The anti-HBsAg antibody 2 is composed of an anti-S protein monoclonal antibody 1, an anti-pre-S2 protein monoclonal antibody and a polyclonal antibody Fab fragment with a molar ratio of 1:0.2:2;

[0043] The polyclonal antibody Fab fragment is prepared and purified by the following method: the polyclonal antibody and the specific Fab processing enzyme are reacted in a 0.2M Tris-HCl buffer system with a pH of 7.2 at a volume ratio of 1:1 at 37°C for 12 hours, Obtain the digested product; pass the digested product through a protein G affinity column to collect the effluent protein P1, fully equilibrate the prot...

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Abstract

The invention relates to a hepatitis B surface antigen chemiluminiscence immune detection reagent kit and application thereof. The reagent kit comprises a magnetic particle reagent indirectly connected with an anti-HBsAg antibody 1, an HBsAg antibody 2 solution marked by a chemiluminiscence marker, a chemiluminiscence substrate solution and a cleaning solution; the anti-HBsAg antibody 1 at least comprises two antibodies or immune reaction fragments of antibodies, and at least comprises an anti-S-protein monoclonal antibody 1, the anti-HBsAg antibody 2 at least comprises two antibodies or immune reaction fragments of the antibodies and at least comprises an anti-S-protein monoclonal antibody 2, the epitopes of HBsAg corresponding to the anti-S-protein monoclonal antibody 1 and the anti-S-protein monoclonal antibody 2 are different, the reagent kit is the first case internationally and has the high sensitivity and accuracy during HBsAg detection, the HBsAg mutant strain detection capability is greatly improved, and the safety for the clinic blood transfusion, operation and the like is remarkably improved.

Description

technical field [0001] The invention relates to a hepatitis B surface antigen chemiluminescence immunoassay kit and application thereof, belonging to the technical field of in vitro detection. Background technique [0002] Hepatitis B surface antigen (HBsAg) is the first serological marker to appear after hepatitis B virus (HBV) infection. The HBV replication cycle is fast and the polymerase is prone to mismatch, and the hepatitis B vaccine and antiviral therapy are widely used. HBsAg mutations are very common in patients with chronic HBV infection. HBsAg mutations can lead to negative HBsAg test results in patients, but HBV DNA still exists in their serum or liver, that is, occult HBV infection (OBI). If OBI patients are used as blood donors or liver transplant donors, they will pose a major risk to the safety of blood transfusion and surgery. At the same time, HBsAg mutation will accelerate the progression of occult hepatitis B to cirrhosis or liver cancer, and trigger H...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/68G01N33/576G01N33/543G01N21/76
CPCG01N21/76G01N33/543G01N33/576G01N33/577G01N33/6854G01N33/5764
Inventor 欧赛英邱春明孟欣沈雪雪
Owner SUZHOU HYBIOME BIOMEDICAL ENG CO LTD
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