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: 2012-12-05
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 anti-HBe antibody-coated luminescent particles

[0088] Preparation:

[0089] 1) Treatment of luminescent particle suspension: a certain amount of luminescent particles is centrifuged in a high-speed refrigerated centrifuge, the supernatant is discarded, a certain amount of MES buffer is added, and the particles are sonicated until the particles are resuspended, and MES buffer is added to adjust the concentration of luminescent particles To 100mg / ml.

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

[0091] 3) MES buffer, 100mg / ml luminescent particle suspension (MES buffer) and 8mg / ml anti-HBe (MES buffer) and mix at a volume ratio of 1:2:5 and mix quickly , Get the reaction solution.

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

Embodiment 2

[0131] Example 2 Preparation of biotin-labeled antibody

[0132] Preparation:

[0133] 1) Antibody treatment: Dialysis anti-HBe to 0.1M NaHCO 3 Solution, measure the antibody concentration and adjust to 1mg / ml.

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

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

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

[0137] 5) Aspirate and transfer the dialyzed biotinylated antibody to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of the biotin-labeled antibody that has passed the quality inspection to 0.5 mg / ml.

[0138] The antibody and Biotin are reacted in different ratios and tested:

[0139] Optical signal detection method:

[0140] Add 2...

Embodiment 3

[0143] Example 3 Preparation of avidin-coated photosensitive particles

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

[0145] Preparation:

[0146] a. Processing 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, sonicate on the ultrasonic cell disruptor until the particles are resuspended, and add MES buffer Adjust the concentration of photosensitive particles to 100mg / ml.

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

[0148] c. Mixing: Mix the processed photosensitive particle suspension, 8mg / ml Avidin and MES buffer at a volume ratio of 2:5:1, and mix quickly to obtain a reaction solution.

[0149] d. Reaction: 25mg / ml NaBH prepared with MES buffer 3 The CN solution was added according to th...

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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 hepatitis B virus e antigen detection particles based on the principle of light-induced chemiluminescence, and preparation and application thereof. Background technique [0002] Immunological detection is a method of detection based on the specific reaction of antigen and antibody. Because it can use isotope, enzyme, chemiluminescent substance, etc. to amplify and display the detected signal, it is often used to detect proteins, hormones, etc. Trace biologically active substances. [0003] my country’s immunological testing has basically undergone radioimmunoassays (which emerged in the 1970s and are still commonly used in hospitals above the county level); enzyme-linked immunoassays (which emerged in the 1980s and are commonly used by clinical institutions); Luminescence is represented by the three stages of photobiological labeling and immunoassay technology (began to be promote...

Claims

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Application Information

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Patent Type & Authority Patents(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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