Kit for detecting habitual abortion pathogenic gene mutation and application thereof

A habitual abortion and pathogenic gene technology, applied in the field of kits for detecting whether habitual abortion pathogenic genes are mutated, can solve problems such as reducing MTHFR activity, blocking the process of homocysteine ​​clearance, and promoting thrombosis tendency. , to reduce the incidence of

Inactive Publication Date: 2011-11-02
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

[0032] A kit for detecting whether there is a mutation in the gene causing recurrent miscarriage for one person, including the following components:

[0033](1) A pair of specific primers and a pair of specific fluorescent probes for detecting the polymorphic genotype of SNP rs769214 on the CTLA4 gene, each primer and probe each 1 OD;

[0034] (2) A pair of specific primers and a pair of specific fluorescent probes for detecting polymorphic genotypes of SNPs rs1800629 and rs1799724 on the TNF-α gene, 1 OD for each primer and probe;

[0035] (3) A pair of specific primers and a pair of specific fluorescent probes for detecting the polymorphic genotype of SNP rs1800796 on the IL-6 gene, 1 OD for each primer and probe;

[0036] (4) A pair of specific primers and a pair of specific fluorescent probes for detecting the polymorphic genotypes of SNPs rs1801133 and rs1801131 on the MTHFR gene, 1 OD for each primer and probe;

[0037] (5) Fluorescent quantitative PCR reaction reagents...

Embodiment 2

[0040] The steps of using a kit for detecting whether a mutation in a habitual abortion gene is used for one person include:

[0041] (1) extract the genomic DNA of the sample;

[0042] (2) Fluorescent quantitative PCR reaction:

[0043] Using the detection kit, carry out 6 independent fluorescent quantitative PCR reactions, each reaction system has a total volume of 10 μl, containing 2 μl of sample genomic DNA template at a concentration of 20ng / μl, 1 μl 10X fluorescent quantitative PCR reaction buffer, 0.1 μl 25mM dNTP mixture, 0.6 μl 25 mM MgCl2 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 , 5.325 μl of deionized water.

[0044] 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 3

[0049] The service of detecting the genetic susceptibility of individual habitual abortion by using the kit for detecting the mutation of the pathogenic gene of habitual abortion

[0050] (1) Instruct the examinee to use the oral sampling swab to sample the oral epithelial cells, and use the silica gel adsorption method to extract the DNA of the oral epithelial cells;

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PUM

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Abstract

The invention discloses a kit for detecting whether relative SNP sites on habitual abortion pathogenic genes mutate or not. The kit provided by the invention mainly comprises the following components of: (1) specific primer pairs and specific fluorescent probe pairs for detecting the genotype of polymorphism of No.rs769214 SNP site on CTLA4 gene, No. rs1800629 and No. rs1799724 SNP sites on TNF-alpha gene, No. rs1800796 SNP site on IL-6 gene, No. rs1801133 and No. rs1801131 SNP sites on MTHFR gene; (2) a routine fluorescence quantitative PCR reaction reagent. The kit provided by the invention is used for detecting the mutant of habitual abortion pathogenic genes so as to rapidly and conveniently check out pathogenic gene carriers and patients, and can be applied in antenatal diagnosis and timely treatment so as to reduce the incidence of habitual abortion and improve pre-natal and post-natal care.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a kit for detecting whether a mutation occurs in a habitual abortion pathogenic gene. 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 natural factors is called spontaneous abortion (Spontaneousabortion, miscarrage). The spontaneous abortion rate accounts for 10% to 15% of all pregnancies, of which 80% are early abortions. [0003] Habitual abortion refers to three or more consecutive spontaneous abortions. Its incidence rate accounts for about 1% of the total number of pregnancies and 15% of the number of spontaneous abortions. The etiolo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 王贻锘
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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