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Hepatitis B virus gene parting method

A technology of hepatitis B virus and genotyping method, which is applied in the directions of biochemical equipment and methods, and the determination/inspection of microorganisms to achieve the effects of reducing missed detection of typing, reducing uneven renaturation and high detection efficiency

Inactive Publication Date: 2005-02-23
THE FIRST PEOPLES HOSPITAL OF FOSHAN +1
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  • Abstract
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Problems solved by technology

However, there is no corresponding public report on the application of this technology in the field of hepatitis B virus genotyping

Method used

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Embodiment Construction

[0022] The outstanding substantive features and positive effects of the present invention can be manifested from the following examples, but they are not intended to limit the present invention.

[0023] Example.

[0024] 1. Specimen source: 300 samples were randomly selected from the 10,000 serum samples of patients who were positive for HBV-DNA in the First People's Hospital of Foshan City as the testing objects for typing. Fifty samples of healthy control serum were from voluntary blood donors, all of which were negative for HBV DNA by PCR.

[0025] 2. Specimen processing: storage, divide the serum and place it at -80°C; for HBV DNA extraction, take 50 μl of serum to be tested, add 50 μl of DNA extraction solution, mix well, boil at 100°C for 10 minutes, and centrifuge at 12,000 r / min for 5 minutes , take the supernatant for later use.

[0026] 3. Main reagents: Biodyne C membrane is the product of PALL company in the United States, EDC [1-ethyl-3-(3-dimethylaminopropyl) ...

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Abstract

A hepatitis B virus gene grouping method by retroactive point hybridizing technology includes: 1) taking X zone sequence of HBV gene group as main designing grouping primmer and probe; 2) fixing active amino-marked probe on nylon membrane to obtain detecting membranous strip; 3) HBV DNA amplifying marked biotin of primmer; 4) amplified product hybridizing with detecting membranous strip, and color developing POD and TMB; 5) discriminating gene grouping end. It achieves low cost and simple operation.

Description

1. Technical field [0001] The invention belongs to a hepatitis B virus genotyping method, in particular to a hepatitis B virus genotyping method established by reverse dot hybridization technology. 2. Background technology [0002] Hepatitis B virus (HBV) infection is globally distributed, with more than 350 million virus carriers, and is a major risk factor for acute, fulminant, and chronic hepatitis, as well as cirrhosis, liver failure, and primary liver cancer. The HBV genome has strong variability. At present, according to the heterology of the nucleotide sequence of the whole HBV gene ≥ 8%, different virus strains can be divided into A ~ H8 genotypes. Different genotypes of the virus may be related to regional distribution, route of infection, pathogenicity, disease progression, and response to treatment. Therefore, the genotyping of HBV is conducive to more in-depth research on the epidemiology, etiology and clinical diagnosis and treatment of HBV infection. Although...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 杨光陈姝崔金环司建华谭家驹
Owner THE FIRST PEOPLES HOSPITAL OF FOSHAN
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