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
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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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