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Hepatitis B virus surface antigen detection particles, preparation thereof and use thereof

A hepatitis B and surface antigen technology, applied in the field of hepatitis B virus surface antigen detection particles

Inactive Publication Date: 2010-03-10
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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  • Hepatitis B virus surface antigen detection particles, preparation thereof and use thereof
  • Hepatitis B virus surface antigen detection particles, preparation thereof and use thereof
  • Hepatitis B virus surface antigen detection particles, preparation thereof and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1 Preparation of Surface Antibody-Coated Luminescent Particles

[0081] Preparation:

[0082] 1) Suspension treatment of luminescent particles: absorb a certain amount of 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 concentration of luminescent particles to 100mg / ml.

[0083] 2) Antibody treatment: HBsAb was dialyzed against 0.05M MES buffer at pH 6.0 (hereinafter referred to as MES buffer). After the dialysis, the concentration was measured and adjusted to 8 mg / ml.

[0084] 3) MES buffer, 100mg / ml luminescent particle suspension (MES buffer) and 8mg / ml anti-HBe (MES buffer) and mixed at a volume ratio of 1:2:5, and mixed quickly , to obtain the reaction solution.

[0085] 4) Prepare 25mg / ml NaBH with MES buffer 3 CN solution was added at a volume ratio of 1:25 to the reaction solut...

Embodiment 2

[0123] Example 2 Preparation of biotin-labeled antibody

[0124] Preparation:

[0125] 1) Antibody treatment: dialyze HBsAb in 0.1M NaHCO 3 solution, the antibody concentration was measured and adjusted to 1 mg / ml.

[0126] 2) Prepare 16.17mg / ml Biotin solution with DMSO.

[0127] 3) Labeling: Take the processed 1 mg / ml HBsAb labeled antibody and the prepared Biotin solution, mix them according to the volume ratio of 72:10000, and mix them uniformly quickly. Stand at 4°C for 12-16 hours.

[0128] 4) Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.00).

[0129] 5) Aspirate the dialyzed biotinylated antibody and transfer it to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of qualified biotin-labeled antibody to 0.5mg / ml.

[0130] React antibodies with Biotin in different ratios and detect:

[0131] Optical signal detection method:

[0132] 25 μl of samples we...

Embodiment 3

[0136] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin

[0137]Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)

[0138] Preparation:

[0139] 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.

[0140] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.

[0141] 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.

[0142] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN so...

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Abstract

The invention relates to a reagent for the diagnosis of hepatitis B and discloses hepatitis B surface antigen detection particles, which are luminous particles coated by a hepatitis B surface antigen.The invention also discloses the preparation and use of the hepatitis B surface antigen detection particles. In addition, the invention also discloses an in-vitro diagnostic kit for determining the hepatitis B surface antigen in a sample and also discloses the using method of the kit. The kit of the invention can be used in combination with other serums and clinic information to diagnose the acute or chronic hepatitis B infection condition of an individual and can also be used for screening hepatitis B in perinatal females to judge the hepatitis B infection risk of the newborns.

Description

technical field [0001] The invention relates to a diagnostic reagent for hepatitis B, in particular to a detection particle for hepatitis B virus surface antigen (HBsAg) 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/52G01N33/543G01N21/64G01N21/76
Inventor 张伟吴旸王海蛟赵卫国
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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