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Habitual abortion genetic detection kit and use thereof

A habitual abortion and kit technology, applied in the direction of measuring devices, microbial determination/inspection, instruments, etc., can solve problems such as blocked homocysteine ​​clearance process, promotion of thrombosis tendency, and increased risk of habitual abortion

Inactive Publication Date: 2009-07-01
XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have shown that the risk genotypes carrying these two MTHFR polymorphisms will reduce the activity of MTHFR, resulting in the blockage of folic acid metabolism and homocysteine ​​clearance process, promoting the tendency of thrombosis, and increasing the incidence of habitual abortion. risk of occurrence

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Use of detection kits

[0030] Step 1: Extraction of DNA template

[0031] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0032] Step 2: Real-time quantitative PCR reaction

[0033] Use the detection kit to carry out 6 independent fluorescent quantitative PCR reactions. The total volume of each reaction is 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10X fluorescent quantitative PCR reaction buffer, and 0.1 μl of 25 mM dNTP mixture. solution, 0.6μl 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM sense primer and antisense primer, 0.25 μl each of 10 μM fluorescent probe with VIC and fluorescent probe with FAM, and 5.325 μl of deionized water.

[0034] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° C. for 1 minute. ...

Embodiment 2

[0038] Example 2. Application of detection kits for the detection of genetic susceptibility of individual habitual abortion

[0039] Step 1: DNA Extraction

[0040] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.

[0041] Step 2: Genotyping Assays

[0042] Using the kit provided by the present invention, the rs769214 SNP site on the CTLA4 gene, the rs1800629 and rs1799724 SNP site on the TNF-α gene, the rs1800796 SNP site on the IL-6 gene, the MTHFR The SNP sites rs1801133 and rs1801131 on the gene were detected by real-time quantitative PCR to determine the genotypes of these 6 SNPs sites.

[0043] Step 3: Analysis of Individual Genetic Susceptibility to Recurrent Miscarriage

[0044] Through the analysis of the SNPs genotypes of the tested subjects, an analysis report sheet of the genetic susceptibi...

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PUM

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Abstract

The invention discloses a reagent kit for detecting personal habitual abortion genetic susceptibility and an application method thereof. The reagent kit comprises six specific primer couples of single nucleotide polymorphic loci genetic type and specific fluorescence probe couples for detecting the associated antigen 4-gene (CTLA4) of cytotoxicity T lymphocyte, tumor necrosis factor-alpha gene (TNF-alpha), interleukin-6 gene (IL-6), and 5, 10-methylene tetrahydrofolate (MTHFR), and general components for fluorescence quantitative PCR detection. The reagent kit can be used to evaluate personal habitual abortion genetic susceptibility.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine, more specifically, the present invention relates to a kit for detecting the genetic susceptibility of individual habitual abortion, by simultaneously detecting the cytotoxic T lymphocyte-associated antigen 4 gene closely related to habitual abortion (CTLA4), tumor necrosis factor-α gene (TNF-α), interleukin-6 gene (IL-6), 5,10-methylenetetrahydrofolate reductase gene (MTHFR) SNP The genotype of morphological loci was used to assess the genetic susceptibility of individuals to recurrent miscarriage. Background technique [0002] If the pregnancy is less than 28 weeks and the fetal weight is less than 1000g and the termination is called abortion (Abortion). Those who terminated before 12 weeks of pregnancy were called early abortion, and those who terminated before 12 weeks to less than 28 weeks of pregnancy were called late abortion. Among them, the abortion caused by natur...

Claims

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Application Information

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IPC IPC(8): G01N21/00C12Q1/68
Inventor 冯哲民邹祖烨
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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