Genome for molecular typing of medulloblastoma and application thereof
A medulloblastoma, molecular typing technology, applied in recombinant DNA technology, microbial determination/examination, DNA/RNA fragment, etc. question
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Embodiment 1
[0099] Training set sample collection and processing:
[0100] The present invention first analyzes the clinical data of 103 patients with medulloblastoma and the gene chip detection data of their biological samples, including relevant clinical data of 8 cases of WNT type, 33 cases of SHH type, 27 cases of Group3 type and 35 cases of Group4 type. Data and expression abundance data of 20,250 genes were used to construct a medulloblastoma gene expression database.
[0101] Screening of 13 specific genes:
[0102] According to the measured value of gene expression abundance, the inventors selected 13 genes closely related to medulloblastoma type from 20,250 genes by means of statistical analysis method variance analysis. These genes were differentially expressed in different subtypes of medulloblastoma, which was statistically significant (P value < 0.0001), see Table 2.
[0103] Table 2: 13 Gene Sets
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[0105] 13 genes based on the construction of statistical analy...
Embodiment 2
[0108] High-throughput sequencing data verification:
[0109] In this example, the inventors analyzed the high-throughput sequencing data of 73 cases of medulloblastoma, including 8 cases of WNT type, 10 cases of SHH type, 16 cases of Group3 type and 39 cases of Group4 type. The 13-gene statistical analysis model was used to distinguish the subtype of each sample, and compared with the clinical diagnosis results, the diagnostic coincidence rate was 75.3%. Taking the SHH type as a reference, the sensitivity of typing was 100.0%, and the specificity was 93.7%. See Table 3 and Table 4 for details.
[0110] Table 3: Classification results of the 13-gene model in 73 cases of high-throughput sequencing data
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[0112] Table 4: Performance indicators of the 13-gene model in 73 cases of high-throughput sequencing data
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Embodiment 3
[0115] In this example, the inventors conducted molecular typing experiments on 285 paraffin tissue samples of medulloblastoma, including 51 cases of SHH type, 46 cases of Group3 type and 188 cases of Group4 type. Experimenters collected medulloblastoma-enriched regions from formalin-fixed, paraffin-embedded tissue blocks by hand scraping, and extracted total RNA; treated with DNase to ensure complete removal of genomic DNA contamination; reverse-transcribed After obtaining cDNA, real-time quantitative polymerase chain reaction (real-timequantitative polymerase chain reaction, RTQ-PCR) of 13 genes was carried out to detect the gene expression level in paraffin-embedded tumor tissue, and the analysis model was used to calculate the relationship between the sample and medulloblastoma subtype. similarity score.
[0116] The subtypes of each sample were discriminated by the 13-gene statistical analysis model, and compared with the clinical diagnosis results, the diagnostic coincid...
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