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Hepatitis b virus surface antigen chemiluminescence immune assay determination kit and method for preparing same

A hepatitis B virus, chemiluminescent immune technology, applied in the field of immunoanalysis medicine, can solve the problems of limited popularization, demand, and inability to be widely used in clinical diagnosis and scientific research work, so as to improve sensitivity, facilitate operation and production, and ensure The effect of sensitivity

Inactive Publication Date: 2009-02-11
北京科美东雅生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, the chemiluminescence immunoassay kit in the prior art is a closed fully automatic chemiluminescence measurement system, which requires an expensive fully automatic chemiluminescence measuring instrument, thus limiting its popularization and use, and cannot be widely used in clinical diagnosis and scientific research Work

Method used

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  • Hepatitis b virus surface antigen chemiluminescence immune assay determination kit and method for preparing same
  • Hepatitis b virus surface antigen chemiluminescence immune assay determination kit and method for preparing same

Examples

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Embodiment 1

[0037]Example 1 Preparation of Hepatitis B virus surface antigen chemiluminescence immunoassay assay kit of the present invention

[0038] 1. Preparation of enzyme-labeled antibody and biotin-conjugated antibody

[0039] 1. The polyclonal antibody or monoclonal antibody of hepatitis B virus surface antibody is coupled with horseradish peroxidase by glutaraldehyde method, fully dialyzed against PBS, added with an equal volume of glycerol, and stored below -20°C for later use.

[0040] 2. Preparation of biotin-conjugated hepatitis B virus surface antibody polyclonal antibody or monoclonal antibody

[0041] (1) Dissolve biotin (BNHS) in N, N-dimethylformamide (DMF) by conventional methods to make 1 mg / mL;

[0042] (2) Use 0.1mol / L NaHCO with a pH value of 9.0 3 Dilute the purified polyclonal antibody or monoclonal antibody of hepatitis B virus surface antibody to 1-2 mg / mL;

[0043] (3) Mix according to the volume ratio of BNHS and antibody is about 1:8 or the weight ratio is ...

Embodiment 2～3

[0098] Examples 2-3 Preparation of Hepatitis B virus surface antigen chemiluminescence immunoassay assay kit of the present invention

[0099] Except that the surface antibody of hepatitis B virus was labeled with alkaline phosphatase, AMPPD was used as the chemiluminescent substrate, plastic beads were used as the carrier, the pH value of the coating solution was 4.5, and the pH value of the blocking solution was 7.5, the rest were prepared with the same The hepatitis B virus surface antigen chemiluminescence immunoassay assay kit was prepared by the same method as in Example 1.

Embodiment 4

[0100] Example 4 Preparation of Hepatitis B virus surface antigen chemiluminescence immunoassay assay kit of the present invention

[0101] Except for using magnetic particles as the carrier, the HBsAg chemiluminescent immunoassay assay kit was prepared in the same manner as in Example 1.

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Abstract

The invention particularly relates to a kit for determining the surface antigen of the hepatitis B virus, and a preparation method thereof, which belongs to the medical field of immunological analysis. The kit comprises (1) a hepatits B virus surface antigen calibration material; (2) an avidin-coated carrier; (3) the polyclonal antibody or monoclonal antibody of a biotinylated hepatitis B virus surface antibody; (4) the polyclonal antibody or monoclonal antibody enzyme-labeled of the surface antibody; and (5) a chemiluminescent primer. Furthermore, the preparation method comprises the following steps of (1) preparing a calibration material from the pure surface antigen; (2) coating the carrier with avidin; (3) performing biotinylation of the polyclonal antibody or monoclonal antibody of the surface antibody; labeling the polyclonal antibody or monoclonal antibody of the surface body with an enzyme; (4) sub-packaging the calibration material and the chemiluminescent primer; and (5) assembling. The kit has the advantages of simpliness, rapidness, sensitiveness, stability and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a hepatitis B virus surface antigen (HBsAg) biotin-avidin immunomagnification technology combined with chemiluminescence immunoassay technology assay kit and a preparation method thereof. Background technique [0002] Hepatitis B virus (HBV) is the pathogen of hepatitis B, and HBV surface antigen is an important serological marker of HBV. The presence of hepatitis B virus surface antigen in serum (or other body fluids) indicates that the subject is infected with hepatitis B virus. The laboratory detection method of hepatitis B virus surface antigen is mainly immunoassay, and the commonly used detection methods are radioimmunoassay (RIA) and enzyme immunoassay (EIA), and the most widely used one is enzyme immunoassay (EIA). [0003] In the past ten years, the research and application of marker immunoassay technology has developed rapidly, and has been wi...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/569G01N33/543G01N21/76
Inventor 杨俊锋胡国茂应希堂
Owner 北京科美东雅生物技术有限公司
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