Method, kit and application for detecting STR locuses of hepatitis B infected related gene

A kit and gene technology, applied in the direction of measuring devices, analysis by making materials undergo chemical reactions, instruments, etc., can solve the problems that there are no research reports on HBV infection

Inactive Publication Date: 2011-09-21
GUIYANG MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The two microsatellite repeat sequences IL-10.G and IL-10.R of the IL-10 gene have been reported to be related to renal transplant rejection and systemic lupus erythematosus, but there is no research report related to HBV infection

Method used

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  • Method, kit and application for detecting STR locuses of hepatitis B infected related gene
  • Method, kit and application for detecting STR locuses of hepatitis B infected related gene
  • Method, kit and application for detecting STR locuses of hepatitis B infected related gene

Examples

Experimental program
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Effect test

Embodiment 1

[0052] Embodiment 1: adopt STRs-PCR method, detect the genotype of two STR sites (IL-10.G, IL-10.R) of the IL-10 gene relevant with hepatitis B virus infection according to the following steps:

[0053] (1) Template DNA preparation:

[0054] Blood samples were taken, and the red blood cell suspension was subjected to phenol (Tris-saturated phenol)-chloroform extraction of genomic DNA, then dissolved in TE buffer (composed of 10mmol / L Tris-HCL (pH 8.0) and 1mmol / L EDTA (pH 8.0)) , DU640 Nucleic Acid Quantitative Apparatus to measure, calculate the content of DNA, and detect the purity of DNA samples at the same time, dilute each sample into an application solution of about 100ng / μl.

[0055] (2) Primer design for two STR sites of IL-10.G and IL-10.R:

[0056] According to the human IL-10 DNA sequence reported by Genbank, the Primer Premier5 software was used to design the primer sequence of the IL-10.G site and the primer sequence for amplifying the IL-10.R site was selected b...

Embodiment 2

[0072] Embodiment 2: adopt STRs-PCR method, detect the genotype of two STR sites (IL-10.G, IL-10.R) of the IL-10 gene relevant with hepatitis B virus infection according to the following steps:

[0073] (1) Template DNA preparation:

[0074] Blood samples were taken, and the red blood cell suspension was extracted with phenol (Tris-saturated phenol) chloroform to extract genomic DNA, then dissolved in TE buffer (composed of 10mmol / L Tris-HCL (pH 8.0) and 1mmol / L EDTA (pH 8.0)), DU640 Nucleic Acid Quantitative Analyzer, dilute each sample into an application solution of about 100ng / μl.

[0075] (2) Primer design for two STR sites of IL-10.G and IL-10.R:

[0076] Use the Primer Premier5 software to design the IL-10.G site primer sequence, refer to the relevant literature to select the primer sequence for amplifying the IL-10.R site, and use fluorescein HEX and 6-FAM at the 5' end of the positive strand primer respectively Marked and sent to Shanghai Jikang Biological Co., Ltd....

Embodiment 3

[0092] Embodiment 3: adopt STRs-PCR method, detect the genotype of two STR sites (IL-10.G, IL-10.R) of the IL-10 gene relevant with hepatitis B virus infection according to the following steps:

[0093] (1) Template DNA preparation:

[0094] Blood samples were taken, and the red blood cell suspension was subjected to phenol-chloroform extraction of genomic DNA, then dissolved in TE buffer to prepare template DNA.

[0095] (2) Primer design for two STR sites of IL-10.G and IL-10.R:

[0096] Use the Primer Premier5 software and refer to the literature to design the primer sequences of IL-10.G and IL-10.R sites:

[0097] Primer sequences for PCR amplification of IL-10.G and IL-10.R microsatellites

[0098]

[0099] (3) Assembly of PCR amplification system: 10×PCR buffer, MgCl 2 (25mmol / L), dNTPs (2.5mol / L), upstream primer G / R (10.0μmol / L), downstream primer G / R (10.0μmol / L), Taq DNA polymerase (5U / μl), template DNA and deionized water

[0100] (4) Mix the reaction system...

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Abstract

The invention discloses a method, a kit and application for detecting genotypes of two STR locuses of a hepatitis B virus infected related IL-10 gene. The method is STRs-PCR, which comprises the steps of preparation of a template DNA, primer design of two STR locuses, assembly of a PCR amplification system, PCR circulation reaction, capillary electrophoresis, fluorescence detection and analysis of a PCR product. Compared with the prior art, the invention ensures that the two STR locuses of the IL-10 gene are related with the susceptibility of hepatitis B virus infection, determines IL-10.G and IL-10.R as novel related genes of the hepatitis B virus infection, detects the IL-10.G and the IL-10.R of the IL-10 genes by adopting the STRs-PCR method, thereby determining the susceptibility of individuals to the hepatitis B, and providing new genetic counseling contents for the prevention and control of hepatitis B virus infected groups.

Description

technical field [0001] The invention relates to a method for detecting the genotypes of two STR loci of IL-10 gene related to hepatitis B virus infection, the kit used and the application in the prevention and control of people infected with hepatitis B virus and genetic counseling, belonging to hepatitis B The field of genetic testing for virus susceptibility. Background technique [0002] According to the research on the characteristics of hepatitis B virus (HBV) infection and pathogenesis, the occurrence and development of hepatitis B are related to the genetic factors of the host, and have a certain influence on the curative effect and prognosis of hepatitis B prevention and treatment. The genetic susceptibility of the host to HBV infection is HBV infection plays an important role in both pathogenesis and disease progression. Studying the host genetic susceptibility of hepatitis B is of great significance. It can not only provide key clues for us to understand the racia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/76G01N27/447
Inventor 单可人王婵娟何燕
Owner GUIYANG MEDICAL UNIVERSITY
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