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ELISA measuring reagent kit for detecting hepatitis B virus kernel antigen in blood serum

A technology of hepatitis B core antibody and core antigen, which is applied in the field of kits, can solve the problems of reducing the sensitivity of HBcAg and application obstacles of detection methods, and achieve the effects of improving specificity and sensitivity, strong practicability, and avoiding interference

Active Publication Date: 2008-03-12
SHANGHAI FOSUN PHARMA (GROUP) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The principle of the above two methods is to first immobilize the hepatitis B virus Danse particles on the microporous plate, and then make it cleavage to release HBcAg, so as to realize the detection of HBcAg, but in the actual detection process, due to the presence of hepatitis B virus in some The serum of patients with viral replication contains very high concentrations of hepatitis B subviral particles. These subviral particles compete with Dansleigh particles to bind to the hepatitis B surface antibody on the microwell plate, so to a certain extent, they will inhibit the interaction between Dansler particles and the microwell plate. Combined, the sensitivity of HBcAg detection is finally reduced, which becomes an obstacle to the application of this type of detection method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0078] Example 1, Preparation of Monoclonal Hepatitis B Core Antibody Pre-coated Microwell Plate

[0079] 1. Coating

[0080] Na 2 CO 3 1.6g

[0081] NaHCO 3 2.9g

[0082] Deionized water 1000ml

[0083] Adjust the pH to 9.6, add an appropriate amount of monoclonal hepatitis B core antibody, mix well and add 100ul per well to each well of the microplate, place it overnight at 4°C, then shake off the liquid in the well and pat dry.

[0084] 2. Washing

[0085] NaHPO 4 .12H 2 O 3.72g

[0086] K H 2 PO 4 0.43g

[0087] NaCl 6.8g

[0088] Tween-20 0.5ml

[0089] Deionized water 1000ml

[0090] Adjust the pH to 7.2, add 150ul per well to each well of the microwell plate, shake it off after standing for 10 seconds, and pat dry to remove the residual monoclonal hepatitis B core antibody that is not combined with the microwell plate.

[0091] 3. Closed

[0092] Bovine Serum Albumin 10g

[0093] NaHPO 4 .12H 2 O 3.72g ...

example 2

[0100] Example 2, preparation of sample treatment agent

[0101] Sample treatment agent ①:

[0102] SDS 10g

[0103] Tween-60 2ml

[0104] Mercaptoethanol 1ml

[0105] NaH 2 PO 4 .2H 2 O 3.8g

[0106] Na 2 HPO 4 10.2g

[0107] Deionized water 1000ml

[0108] Sample treatment agent②:

[0109] SDS 1g

[0110] Tween-60 0.5ml

[0111] Mercaptoethanol 5ml

[0112] NaH 2 PO 4 .2H 2 O 1.9g

[0113] Na 2 HPO 4 5.1g

[0114] Deionized water 1000ml

[0115] Sample treatment agent③:

[0116] SDS 5g

[0117] Tween-60 1ml

[0118] Mercaptoethanol 5ml

[0119] NaH 2 PO 4 .2H 2 O 1.9g

[0120] Na 2 HPO 4 5.1g

[0121] Deionized water 1000ml

[0122] After the sample processing agent is prepared, it is stored in a 4°C environment.

example 3

[0123] Example 3, preparation of enzyme-labeled antibody working solution

[0124] Bovine Serum Albumin 5g

[0125] NaHPO 4 .12H 2 O 3.72g

[0126] K H 2 PO 4 0.43g

[0127] NaCl 6.8g

[0128] Deionized water 1000ml

[0129] Adjust the pH to 7.2, add an appropriate amount of horseradish peroxidase-labeled polyclonal hepatitis B core antibody, and store at 4°C.

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Abstract

The present invention discloses an enzyme association immunity test reagent kit and its operation instruction to test hepatitis B core antigen. Wherein, the reagent kit mainly comprises a monoclonal hepatitis B core antigen prepackaged micro-perforated plate, sample processing agent and polyclone hepatitis B core antigens marked with horse radish peroxidase. During operation, a sample to be tested is firstly preprocessed with the sample processing agent and then added into the monoclonal hepatitis B core antigen prepackaged micro-perforated plate. And then, polyclone hepatitis B core antigens marked with horse radish peroxidase and zymolytes are added to colorize. Stop solution is added to stop reaction and finally the measuring result is confirmed through color comparison. The present invention can detect hepatitis B core antigen in blood serum, ensure quite high conformance ratio in aspect of HBV-DNA detecting result, achieve requirements of clinical examination, have characteristics of convenience, sensitivity and stability, supplement conventional hepatitis B virus detecting method and bring better clinical operation.

Description

technical field [0001] The invention relates to a kit, in particular to an enzyme-linked immunoassay kit for detecting the core antigen of hepatitis B virus in serum. Background technique [0002] There are three forms of virus particles in the serum of patients with hepatitis B, the most common ones are small spherical particles with a diameter of about 17-25nm, and the slightly lesser ones are tubular or filamentous particles with a diameter of about 20-22nm and different lengths Particles, these two kinds of particles belong to the subviral structure, both of which are excess virus shells, only contain HBV surface antigen but not HBV DNA, so they are non-infectious, and there is another kind of virus particles with a diameter of about 42nm, and the outer layer is the surface of HBV The envelope composed of antigen (HBsAg), and the inner layer is the nucleocapsid composed of HBV core antigen (HBcAg), which wraps the HBV genome and related polymerase. This type of virus par...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/576
Inventor 龚育平
Owner SHANGHAI FOSUN PHARMA (GROUP) CO LTD
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