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Non-invasive detection kit for susceptibility gene of basal cell carcinoma

A basal cell carcinoma, detection kit technology, applied in the field of molecular biology, can solve the problems of cell proliferation, DNA entry, cell cycle arrest affecting the transcriptional activation function of P53 gene, etc.

Inactive Publication Date: 2013-10-30
解码(上海)生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If p53 function is inactivated or degraded, it will cause damaged DNA to enter the cell cycle, and the cells will proliferate unrestrictedly, leading to the occurrence of skin basal cell carcinoma
Studies have found that the P53 Arg72Pro polymorphism affects the transcriptional activation function of the P53 gene, the arrest of the cell cycle, etc., causing unlimited cell proliferation and eventually leading to the occurrence of skin basal cell carcinoma

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Use of detection kits

[0025] 1. Extract DNA template

[0026] The epithelial cells of the oral mucosa of the subjects were scraped, and the genomic DNA was extracted by the phenol-chloroform method.

[0027] 2. PCR amplification reaction

[0028] Use the PCR reaction component in the detection kit, which contains the following primer pairs:

[0029] (1) Mn-SOD (Ala-9Val) forward primer: 5'-CAGCCCCAGCCTGCGTAGACG-3'

[0030] Mn-SOD (Ala-9Val) reverse primer: 5'-GCGTTGATGTGAGGTTCCAG-3'

[0031] (2) P53 (Arg72Pro) forward primer: 5'- CGTTCTGGTAAGGACAAGGGTT-3'

[0032] P53 (Arg72Pro) reverse primer: 5'- AAGAAGCCCAGACGGAAACC-3'

[0033] (3) XRCC1 (Arg399Gln) forward primer: 5'- TGACCTTGCGGGACCTTAG-3'

[0034] XRCC1 (Arg399Gln) reverse primer: 5'-CCAAGACCCTTTTCACTCCTATC-3'

[0035] The reaction system for PCR amplification is: 10×PCR reaction buffer 2.5ml; 25mM dNTP mixture 0.2ml, 5U / ul Taq enzyme 0.125ml, DNA template 1ml (about 12-15ng), 20uM forward...

Embodiment 2

[0054] Example 2. Gene non-invasive testing services for screening people at risk of skin basal cell carcinoma

[0055] 1. Non-invasive sampling

[0056] The physician in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral mucosal epithelial cells, and the samples were stored at room temperature with cell preservation solution.

[0057] 2. DNA extraction

[0058] DNA was extracted from the collected oral mucosal cells by the phenol-chloroform method.

[0059] 3. Genotype detection

[0060] Using the kit provided by the present invention, the single nucleotide polymorphism site Ala-9Val (rs1799725) on the Mn-SOD gene and the single nucleotide polymorphism site Arg72Pro on the P53 gene of the subject's whole genome DNA (rs1042522), the single nucleotide polymorphism site Arg399Gln (rs1799782) on the base of XRCC1 were sequenced separately to determine the genotypes of these three SNPs sites.

[0061] 4. Bioinformatics analysis a...

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Abstract

The invention provides a non-invasive detection kit for susceptibility gene of basal cell carcinoma. The kit comprises a specific primer and a DNA sequencing primer which are used for detecting the genotypes of a Mn-SOD gene, a P53 gene and an XRCC1 (X-ray repair cross-complementing group 1) gene, a PCR (Polymerase Chain Reaction) reaction system, a PCR product purification system and a DNA sequencing reaction system, etc. The kit disclosed by the invention is used for evaluating the suffering risk level and degree of basal cell carcinoma in people by detecting the genotypes of three single nucleotide polymorphic sites closely related with occurrence of basal cell carcinoma, and finally guiding Chinese people to pointedly prevent occurrence of basal cell carcinoma from gene dimension according to the gene detection results of a detected person, thereby reducing the morbidity risk rate of basal cell carcinoma. A sample used in the detection kit provided by the invention is a mouth mucosa cell, and a collection method is free of pain and non-invasive, and can prevent cross infection.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to biomedicine and biotechnology, and specifically relates to a skin basal cell carcinoma susceptibility gene detection kit, which evaluates the risk level of skin basal cell carcinoma from the genetic level according to the detection results, and serves as a preventive and Directions for the treatment of basal cell carcinoma of the skin. Background technique [0002] Skin basal cell carcinoma is a common low-grade malignant tumor. In my country, skin cancer accounts for about 1.5% of all malignant tumors, and the incidence of skin basal cell carcinoma (SCC) and squamous cell carcinoma (BCC) account for More than 90% of cancer, and it is increasing year by year. Patients with basal cell carcinoma of the skin are more prone to regional lymph node metastasis, and its risk is much higher than that of squamous cell carcinoma. Therefore, in recent years, there have been more and more researche...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 潘加奎郑俊斌
Owner 解码(上海)生物医药科技有限公司
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