Fluorescent quantitative PCR detection system applied to melanoma gene screening and application of detection system

A fluorescence quantitative, melanoma technology, applied in the field of biological detection, can solve problems such as weak site correlation, and achieve the effect of changing living habits, strong specificity, and avoiding inducing factors

Inactive Publication Date: 2018-04-10
SHENZHEN MEIGENE MEDICAL LAB
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Problems solved by technology

Most of the existing studies on melanoma susceptibility genes are based on the strong association loci obtained from the large-sample GWAS study of the European population. When applied to the Asian population, espec

Method used

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  • Fluorescent quantitative PCR detection system applied to melanoma gene screening and application of detection system
  • Fluorescent quantitative PCR detection system applied to melanoma gene screening and application of detection system
  • Fluorescent quantitative PCR detection system applied to melanoma gene screening and application of detection system

Examples

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Effect test

Embodiment 1

[0056] In this example, the melanoma fluorescent quantitative PCR detection kit was used to detect melanoma-related genes, in which 7 polymorphic sites on 7 genes related to melanoma were detected, including rs401681, rs7023329, rs1847142, rs258322 , rs132985, rs10757257 and rs7412746 sites, a total of 5 sites. Using specifically designed primers and probes, the amplification system and conditions were optimized, and the detection of seven polymorphic sites on seven genes was completed in four systems.

[0057] In this example, a single fluorescently labeled probe, Roche 480 platform, and finally detected rs401681, rs7023329, rs1847142, rs258322, rs132985, rs10757257 and rs7412746 loci by melting curve peak pattern.

[0058] In the specific rs401681, rs7023329, rs1847142, rs258322, rs132985, rs10757257 and rs7412746 site double fluorescent quantitative PCR detection method, for rs401681, rs7023329, rs1847142, rs258322, rs13657257 and rs10757257 primers are designed as follow...

Embodiment 2

[0092] The seven loci in Example 1 are all from articles published in top international journals. The article obtained seven susceptibility loci directly related to melanoma through GWAS research, and calculated the risk alleles of the seven loci Genotype risk value, ie OR value.

[0093] Multiple GWAS studies have found that common genetic variations in TERT-CLPTM1, CDKN2A, TYR, MC1R, PLA2G6, MTAP, and ARNT genes are closely related to the risk of melanoma. Marta et al. found that the T allele of rs401681 on the TERT-CLPTM1 locus and longer telomere length can significantly increase the risk of melanoma; DT Bishop et al. confirmed the rs7023329 of the CDKN2A gene through case-control experiments in two populations , rs258322 on the MC1R gene and rs1847142 on the TYP gene are all significantly associated with the risk of melanoma. Among them, MC1R and TYP are important genes for pigment synthesis, which have been confirmed to be closely related to melanoma risk factors such as...

Embodiment 3

[0117] The method of the present invention extracts DNA by collecting the gene sample of examinee then, detects with Roche LightCycler480 real-time quantitative instrument, the result obtained can refer to figure 1 , 3, 5, 7, 9, 11, 13.

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Abstract

The invention relates to the technical field of biological detection, in particular to a fluorescent quantitative PCR detection system, and further discloses an application of the fluorescent quantitative PCR detection system in detecting melanoma. The fluorescent quantitative PCR detection system comprises a detection kit for specific-site genes; scores of a melanoma risk are calculated in accordance with a risk value of each site on the melanoma risk, population classification is implemented on the basis of an existing database, and the melanoma risk level of a person is obtained in accordance with scores of the person, so that disease risks of detected objects are assessed, life styles of high-risk objects are improved, and meanwhile, induction factors are avoided and physical examination frequency is improved. Meanwhile, a database for Chinese people is established on the basis of seven selected gene polymorphism sites; therefore, a foundation is laid for assessing melanoma in theChinese people.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a fluorescent quantitative PCR detection kit, and further discloses its application for detecting melanoma. Background technique [0002] With the continuous development of scientific research, after the completion of the Human Genome Project and the full development of the Post-Genome Project, the application of genetic engineering-led biotechnology in the field of sports issues has received widespread attention. At present, there has been preliminary progress in the selection of sports materials using the theory and methods of genetics. Commonly used gene detection methods include: direct sequencing (DS), ligase detection reaction (LDR), restriction fragment length polymorphism (RFLP), denaturing high performance liquid chromatography analysis (denaturing high performance liquid chromatography, DHPLC) and quantitative PCR, among which direct sequencing using a DNA...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858C12Q1/6886
CPCC12Q1/6858C12Q1/6886C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/165C12Q2537/143C12Q2563/107
Inventor 梁良周文根李欣姜萍萍黄君任飞詹延延赵世林李政吴少鸿
Owner SHENZHEN MEIGENE MEDICAL LAB
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