Hepatitis B virus e antigen testing corpuscle, preparation and application thereof

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

Active Publication Date: 2008-08-27
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 e antigen testing corpuscle, preparation and application thereof
  • Hepatitis B virus e antigen testing corpuscle, preparation and application thereof
  • Hepatitis B virus e antigen testing corpuscle, preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1 Preparation of luminescent particles coated with anti-HBe antibody

[0088] Preparation:

[0089] 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.

[0090] 2) Antibody treatment: Anti-HBe was dialyzed in 0.05M MES buffer solution with pH 6.0 (hereinafter referred to as MES buffer solution). After the dialysis was completed, the concentration was measured and adjusted to 8 mg / ml.

[0091] 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.

[0092] 4) Prepare 25mg / ml NaBH with MES buffer 3 CN solution was added at a vo...

Embodiment 2

[0131] Example 2 Preparation of biotin-labeled antibody

[0132] Preparation:

[0133] 1) Antibody treatment: dialyze anti-HBe in 0.1M NaHCO 3 solution, the antibody concentration was measured and adjusted to 1 mg / ml.

[0134] 2) Prepare 16.17mg / ml Biotin solution with DMSO.

[0135] 3) Labeling: take the processed 1 mg / ml anti-HBe labeled antibody and the prepared Biotin solution, mix them according to the volume ratio of 10000:54, and mix them uniformly quickly. Stand at 2-8°C for 12-16 hours.

[0136] 4) Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.00).

[0137] 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.

[0138] React antibodies with Biotin in different ratios and detect:

[0139] Optical signal detection method:

[0140] 25 μl of samp...

Embodiment 3

[0143] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin

[0144] Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)

[0145] Preparation:

[0146] 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.

[0147] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.

[0148] 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.

[0149] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN s...

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Abstract

The invention relates to a diagnosing reagent for hepatitis B, disclosing detection grains for e antigens of the hepatitis B virus, which are of the luminous grains coated by anti-HBE antibodies. The invention also discloses preparation and application for the detection grains for e antigens of the hepatitis B virus; moreover, the invention further discloses an outside-body diagnosis reagent box for detecting e antigens of the hepatitis B in a blood serum sample of human beings as well as a method for utilizing the light excitation chemiluminescence principle to quantitatively and qualitatively detect e antigens of the hepatitis B virus. The reagent box of the invention can be jointly used to diagnose the individual acute or chronic hepatitis B together with other blood serums and clinic information, and screen the hepatitis B for women in the perinatal period so as to judge the risk of newborn babies contaminating the hepatitis B.

Description

technical field [0001] The invention relates to a diagnostic reagent for hepatitis B, in particular to a particle for detecting hepatitis B virus e antigen 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, and the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/546G01N21/63
Inventor 王海蛟赵卫国张向辉
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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