Core antibody testing fine particles for hepatitis b virus, and preparation and application thereof

A kind of hepatitis B, core antibody technology, applied in the field of hepatitis B diagnostic reagents

Active Publication Date: 2010-07-07
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the field of clinical detection, there is no light-induced chemiluminescence detection reagent available, especially for tumor markers and hepatitis detection items

Method used

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  • Core antibody testing fine particles for hepatitis b virus, and preparation and application thereof
  • Core antibody testing fine particles for hepatitis b virus, and preparation and application thereof
  • Core antibody testing fine particles for hepatitis b virus, and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1 Preparation of Hepatitis B Core Antigen-Coated Luminescent Particles

[0085] The improved preparation method of the hepatitis B core antigen-coated luminescent particles is as follows:

[0086] 1) Suspension treatment of luminescent particles: absorb a certain amount of carboxyl luminescent particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically break until the particles are resuspended, add MES buffer to adjust the luminescent particles Concentration to 100mg / ml.

[0087] 2) Antigen treatment: HbcAg antigen was dialyzed in 0.05M MES buffer solution with pH 6.0 (hereinafter referred to as MES buffer solution). After the dialysis, the concentration was measured and adjusted to 8mg / ml.

[0088] 3) MES buffer, 100mg / ml of luminescent microparticle suspension (MES buffer) and 8mg / ml of HbcAg (MES buffer) and mixed at a volume ratio of 1:1:1, mixed rapidly to obtain The reactio...

Embodiment 2

[0139] Example 2 Preparation of biotin-labeled core antigen

[0140] Preparation:

[0141] 1) Antigen treatment: dialyze HBcAg against 0.1M NaHCO 3 solution, the antigen concentration was determined and adjusted to 1 mg / ml.

[0142] 2) Prepare 16.17mg / ml Biotin solution with DMSO.

[0143] 3) Labeling: take the processed 1 mg / ml HBcAg labeled antigen and the prepared Biotin solution, mix them according to the volume ratio of 324:10000, and mix them uniformly quickly. Stand at 4°C for 12-16 hours.

[0144] 4) Dialysis: Dialyze the reacted biotin-labeled antigen against biotin-labeled dialysis buffer (pHg.0).

[0145] 5) Aspirate the dialyzed biotinylated antigen and transfer it to a clean centrifuge tube, and take a sample to determine the antigen concentration. Adjust the concentration of qualified biotin-labeled antigen to 0.5mg / ml.

[0146] React the antigen with Biotin in different ratios and detect:

[0147] Optical signal detection method:

[0148] Add 25 μl of samp...

Embodiment 3

[0152] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin

[0153] Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)

[0154] Preparation:

[0155] a. Treatment of photosensitive particle suspension: absorb a certain amount of photosensitive particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically sonicate on the ultrasonic cell disruptor until the particles are resuspended, and then add MES buffer Adjust the photosensitive particle concentration to 100mg / ml.

[0156] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.

[0157] c. Mixing: Mix the treated photosensitive microparticle suspension, 8 mg / ml Avidin and MES buffer at a volume ratio of 2:5:1, and mix quickly to obtain a reaction solution.

[0158] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN s...

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Abstract

The invention relates to hepatitis B diagnosis reagent, which discloses core antibody testing fine particles for hepatitis B, which are luminous fine particles for coating core antibody of hepatitis B. The invention also discloses a preparation and an application for the core antibody testing fine particles for hepatitis B. In addition, the invention further discloses an in vitro diagnosis kit for measuring the core antibody of hepatitis B and simultaneously discloses a method for using the kit. The kit can be used jointly with other blood serum and clinical information to diagnose the condition of acute or chronic hepatitis B infection of an individual, and can also be used for sieving hepatitis B of females in the perinatal period, so as to judge the danger of hepatitis B infection of newly born babies.

Description

technical field [0001] The invention relates to a diagnostic reagent for hepatitis B, in particular to a hepatitis B virus core antibody (HBcAb) detection particle based on the principle of light-induced chemiluminescence, its preparation and application. Background technique [0002] Immunological detection is a means of detection based on the specific reaction of antigens and antibodies. Because it can use isotopes, enzymes, chemiluminescent substances, etc. to amplify and display the detected signals, it is often used to detect proteins, hormones, etc. trace bioactive substances. [0003] Immunological testing in my country has basically experienced radioimmunoassay (emerged in the 1970s, and is still widely used in hospitals above the county level); enzyme-linked immunoassay (emerged in the 1980s, commonly used in various clinical institutions); Photobiological labeling and immunoassay technology represented by luminescence (began to be popularized and used in the 1990s,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/543G01N33/52G01N21/76G01N21/64
Inventor 赵卫国王海蛟陈晨辉
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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