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Reagent and method for in-vitro diagnosis of hepatitis B virus antibody

A hepatitis B virus, in vitro diagnosis technology, applied in the field of immunomedical analysis, can solve problems such as poor repeatability and inaccurate measurement results

Active Publication Date: 2021-09-07
深圳市辰纳生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional ELISA method has the problems of poor repeatability and inaccurate measurement results.

Method used

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  • Reagent and method for in-vitro diagnosis of hepatitis B virus antibody
  • Reagent and method for in-vitro diagnosis of hepatitis B virus antibody
  • Reagent and method for in-vitro diagnosis of hepatitis B virus antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Preparation of hepatitis B virus surface antibody in vitro diagnostic reagents:

[0075] (1) Preparation of solid-phase antigen: coat the antigen on the surface of the solid-phase carrier

[0076] 1) After dissolving 2 mg of HBsAg recombinant protein in 1 L of coating buffer, add it to magnetic microspheres with 5 times the amount of HBsAg recombinant protein, and place at 4°C overnight;

[0077] 2) After washing and purifying, block;

[0078] Wherein, the magnetic microsphere is based on Fe 3 o 4 Nano-magnetic particles are used as the inner core, and polystyrene is used as the outer shell;

[0079] Described coating buffer solution is the Na that concentration is 0.05mol / L, pH is 9.6 2 CO 3 -NaHCO 3 buffer;

[0080] Washing and purifying with physiological saline;

[0081] Bovine serum albumin solution is used as a blocking agent for blocking, and the mass concentration of bovine serum albumin in the bovine serum albumin solution is 1%-5%.

[0082] (2) Prepar...

Embodiment 2

[0098] The method for using the reagent to identify the surface antibody of hepatitis B virus comprises the following steps:

[0099] 1) Sample addition: Mix 1 drop (about 0.05 g) of isolated fresh human plasma with 2 mg of solid-phase antigen and 1 mL of enzyme-labeled antigen, and incubate at 37°C for 30 min to obtain a mixture containing the double-antigen sandwich complex;

[0100] The solid-phase antigen is that the hepatitis B surface antigen recombinant protein is coated with Fc 3 o 4 The product on the surface of magnetic microspheres made of nano-magnetic particles as the core and polystyrene as the shell;

[0101] The enzyme-labeled antigen is a product obtained by labeling the hepatitis B surface antigen recombinant protein with horseradish peroxidase;

[0102] 2) Separation and purification: the mixture is placed in a magnetic separation device, the above-mentioned complex is precipitated under the action of an external magnetic field, the supernatant is removed,...

Embodiment 3

[0105] According to Example 2, after mixing isolated fresh human plasma with solid-phase antigen and enzyme-labeled antigen, a mixture containing a double-antigen sandwich complex is obtained, and the complex needs to be separated and purified, which includes the following steps:

[0106] a) Putting the mixture containing the double-antigen sandwich complex into a test tube to form a mixed system, placing the mixed system in a magnetic field to layer up and down, forming a supernatant layer at the top and a complex precipitation layer at the bottom;

[0107] b) removing the supernatant layer and retaining the complex precipitation layer;

[0108] c) injecting a cleaning solution into the composite precipitation layer to obtain a composite-cleaning solution system;

[0109] Change the direction of the magnetic field, and the complexes in the complex precipitation layer move in the complex-cleaning liquid system to clean the complexes;

[0110] d) After the cleaning is complete...

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Abstract

The invention discloses a reagent for hepatitis B virus antibody in-vitro diagnosis, the reagent comprises a solid phase antigen, an enzyme-labeled antigen and a chromogenic substrate, the solid phase antigen comprises a solid phase carrier and an antigen coated on the surface of the solid phase carrier, the enzyme-labeled antigen comprises an enzyme for labeling the antigen and an enzyme-labeled antigen, the antigen is hepatitis B antigen recombinant protein, the antigens in the solid-phase antigen and the enzyme-labeled antigen are the same hepatitis B antigen recombinant protein, the solid-phase carrier comprises magnetic microspheres, the enzyme comprises horse radish peroxidase, and the chromogenic substrate comprises TMB. The invention also discloses a method for preparing the reagent and a method for identifying the hepatitis B virus antibody by using the reagent. The reagent is used for detecting the hepatitis B virus antibody, has the advantages of good specificity, high sensitivity, accurate result, good repeatability and strong clinical applicability, and can accurately reflect the content index of the hepatitis B virus antibody in a human body.

Description

technical field [0001] The invention relates to the field of immunomedicine analysis, in particular, the invention relates to a reagent and method for in vitro diagnosis of hepatitis B virus antibody. Background technique [0002] At present, the laboratory methods for detecting hepatitis B antibodies in serum mainly include enzyme-linked immunosorbent assay (ELISA), chemiluminescence quantification, and gold standard method, among which chemiluminescence is a more accurate method for detecting hepatitis B antibodies, but it is expensive and difficult to popularize promote. Because ELISA has high sensitivity and specificity and low cost, most hospitals now use ELISA to detect hepatitis B antibody. However, the traditional ELISA method has the problems of poor repeatability and inaccurate measurement results. Contents of the invention [0003] The purpose of the present invention is to provide a reagent and method for in vitro diagnosis of hepatitis B virus antibody, whic...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/535G01N21/76
CPCG01N33/68G01N33/54326G01N33/535G01N21/76
Inventor 汪紫晶于琴丁军发谢钧
Owner 深圳市辰纳生物科技有限公司
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