Hepatits B virus e antibody assay kit and assay method thereof

A hepatitis B and antibody detection technology, applied in the direction of biological testing, measuring devices, material inspection products, etc.

Active Publication Date: 2010-01-06
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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  • Hepatits B virus e antibody assay kit and assay method thereof
  • Hepatits B virus e antibody assay kit and assay method thereof
  • Hepatits B virus e antibody assay kit and assay method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1 Preparation of luminescent particles coated with anti-HBe antibody

[0085] Preparation:

[0086] 1) Treatment of luminescent particle suspension: absorb a certain amount of luminescent particles (particle size 250±20nm) 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 adjusted the concentration of luminescent microparticles to 100mg / ml.

[0087] 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.

[0088] 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.

[0089] 4) Prepare 25mg / ml NaBH with MES buffer 3 C...

Embodiment 2

[0127] Example 2 Preparation of biotin-labeled antibody

[0128] Preparation:

[0129] 1) Antibody treatment: dialyze anti-HBe in 0.1M NaHCO 3solution, the antibody concentration was measured and adjusted to 1 mg / ml.

[0130] 2) Prepare 16.17mg / ml Biotin solution with DMSO.

[0131] 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.

[0132] 4) Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.00).

[0133] 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.

[0134] According to the above preparation method, change the molecular ratio of antibody and Biotin to react and detect:

[0135] Op...

Embodiment 3

[0139] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin

[0140] Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)

[0141] Preparation:

[0142] 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.

[0143] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.

[0144] 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.

[0145] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN s...

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Abstract

The invention relates to a diagnostic reagent for hepatits B, and discloses a hepatits B virus e antibody assay kit and an assay method thereof. The invention adopts light initiated chemiluminescence assay technology, and discloses the hepatits B virus e antibody assay kit, which comprises luminescent particles coated by anti-HBe antibody, anti-HBe antibody marked by biotin and neutralized e antigen. The invention also discloses a method for qualitatively or quantitatively assaying hepatits B e antibody by the light initiated chemiluminescence assay technology. The assay kit can be combined with other serum and clinical information for diagnosing infectious conditions of acute or chronic hepatits B of an individual, and also can be used for screening hepatits B of female in perinatal period to judge the hazard that neonates are infected with the hepatits B. Besides, the kit has the characteristics of high sensitivity, wide assay range and the like, and the assay method has higher sensitivity and better assay range than an enzyme-immunoassay method methodologically.

Description

technical field [0001] The invention relates to a diagnostic kit for hepatitis B, in particular to a detection kit and a detection method for hepatitis B virus e antibody based on the principle of light-induced chemiluminescence. Background technique [0002] Hepatitis B virus (HBV) is endemic and a major cause of liver disease. HBV can be transmitted directly through blood and body fluids. Common modes of transmission include blood transfusions, injections, direct contact with open wounds, sexual intercourse, and mother-to-child transmission at birth. The average incubation period after HBV infection is 6 to 8 weeks (1 to 6 months). Common clinical symptoms include malaise, fever, gastroenteritis, jaundice, which can lead to typical jaundice hepatitis, asymptomatic hepatitis, fulminant hepatitis, and chronic hepatitis. In adults, 90% to 95% of patients with acute HBV infection recover and clear the virus. About 5 to 10% of patients become chronic carriers. Among HBV-in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/52
Inventor 王海蛟赵卫国
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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