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
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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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