Fluorescent quantitative PCR detection system for Wilms' tumor gene screening and application thereof

A Wilms tumor, fluorescence quantitative technology, applied in the fluorescence quantitative PCR detection kit, the application field of Wilms tumor gene detection, can solve the problem of weak locus correlation, change living habits, increase the frequency of physical examination, The effect of improving lifestyle

Inactive Publication Date: 2018-03-23
SHENZHEN MEIGENE MEDICAL LAB
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  • Claims
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Problems solved by technology

[0008] Existing Wilms tumor gene research, these loci are based on the strong association loci obtained from the large-sample GWAS study of the European population. When applied to the Asian population, especially the Chinese population, the locus association is weak. It is difficult to obtain more accurate results through testing, so it is necessary to establish an assessment system for the Chinese population

Method used

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  • Fluorescent quantitative PCR detection system for Wilms' tumor gene screening and application thereof
  • Fluorescent quantitative PCR detection system for Wilms' tumor gene screening and application thereof
  • Fluorescent quantitative PCR detection system for Wilms' tumor gene screening and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0047] In this example, Wilms tumor fluorescent quantitative PCR detection kit was used to detect Wilms tumor-related genes, in which three polymorphic sites on three genes related to Wilms tumor were detected, including rs3755132 , rs807624 and rs790356 sites, a total of 3 sites. Using specifically designed primers and probes, the amplification system and conditions are optimized, and the detection of three polymorphic sites on three genes is completed simultaneously in the same system.

[0048] In this embodiment, a single fluorescently labeled probe is used to implement 480 platform, and finally detected rs3755132, rs807624 and rs790356 sites by melting curve peak pattern.

[0049] Specifically, in the double fluorescent quantitative PCR detection method for rs3755132, rs807624 and rs790356 sites, the design of primers and probes for rs3755132, rs807624 and rs790356 sites is shown in Table 1 below.

[0050] Table 1 Primer and probe design

[0051]

[0052] For the ab...

Embodiment 2

[0073] The three loci in Example 1 are from the articles published by C Turnbull and E Perdeaux, etc. The article obtained three susceptibility loci directly related to Wilms tumor through GWAS research, and calculated the three loci The risk allele type risk value of the point, that is, the OR value. GWAS studies have found that genetic variations in 2p24 and 11q14 genes may be related to the occurrence of Wilms tumor. In order to explore the relationship between genetic variation and Wilms tumor, C Turnbull and E Perdeaux et al. compared and analyzed the genes of 757 Wilms tumor patients and 18.79 million normal people. Two genes, 2p24 and 11q14, are strongly associated with the pathogenesis of Wilms tumor.

[0074] Evaluation of the above 3 loci, the different genotypes and their risk values ​​(OR) are as follows: the OR values ​​of the GG-TG-TT genotype at the 2p24 gene rs3755132, rs807624 and rs790356 loci are 2.19-1.48-1 respectively; The OR values ​​of the TT-GT-GG ge...

Embodiment 3

[0091] The test results of the next subject are shown in Table 6 below.

[0092] The genotype of rs3755132 in the 2p24 region of a subject is TT, and the corresponding OR value is 1; the genotype of rs807624 in the 2p24 region is GT, and the corresponding OR value is 1.33; the genotype of rs790356 in the 2p24 region is AG, and the corresponding OR value is 1.43. The above OR value comes from the literature research of NCBI.

[0093] Calculate the subject's individual osteosarcoma susceptibility value P=1*1.33*1.43=1.9019.

[0094] Table 6 Genetic testing results of the subjects

[0095]

rs3755132

rs807624

rs790356

Subject

TT

GT

AG

HG00403

TT

GT

GG

HG00404

TG

TT

GG

HG00406

TT

TT

GG

[0096] There are 504 East Asians in the 1000 Genomes database. For example, HG00403 has P=1*1.33*2.04 =2.7132, and other people can also be calculated. The average genotype score of the East Asia...

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Abstract

The invention relates to the technical field of biological detection, in particular to a fluorescent quantitative PCR detection kit, and further discloses application of the fluorescent quantitative PCR detection kit in detecting Wilms' tumor. A fluorescent quantitative PCR detection system comprises the detection kit for specific locus gene; a Wilms' tumor risk score is calculated according to aWilms' tumor risk value of each locus, population grading is performed according to an existing database, a Wilms' tumor risk grade of an individual is acquired through each individual score, in thisway, disease risk of a subject is evaluated, and people with high risk are guided to improve living styles, avoid inducing factors and increase physical examination frequency. In addition, by buildinga Chinese population database for three selected gene polymorphic loci, a foundation is laid for evaluating Wilms' tumor unique to Chinese population.

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 detection of nephroblastoma gene. 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 cancer research has attracted widespread attention. At present, there has been preliminary progress in the study of tumor susceptibility 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 seque...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q2600/156C12Q2531/113C12Q2563/107C12Q2537/165
Inventor 吴少鸿周文根李欣姜萍萍黄君任飞詹延延赵世林李政
Owner SHENZHEN MEIGENE MEDICAL LAB
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