NGS-based brain tumor molecular diagnosis analysis method

Abstract

The invention discloses an NGS-based brain tumor molecular diagnosis analysis method. The method comprises the following steps: constructing a cluster based on an Amazon cloud platform; establishing an analysis process of the molecular marker for analyzing the brain tumor on the cluster; a sequence alignment module; removing the repetitive sequence module; a gene fusion analysis module; a call variation module; a cnv analysis structure variation module; the method disclosed by the invention has the advantages of high sensitivity, high specificity, high repeatability, low experimental cost andshort analysis period, and can be used for comprehensively analyzing the hot spot mutation of the TERT promoter, the deletion states of the chromosomes 1p and 19q and other main brain tumor related molecular markers at one time. The NGS-based analytical technique of the present invention enables reliable detection of 1p and / or 19q deletions in the background tumor tissue of 70% normal cells, is more sensitive than microsatellite-based LOH analysis, and requires less DNA.

Description

technical field [0001] The invention relates to the field of molecular diagnosis of brain tumors, in particular to an analysis method for molecular diagnosis of brain tumors based on NGS. Background technique [0002] For the past 100 years, brain tumors have been graded primarily by histomorphology, based on the microscopic similarity of different cells of origin and the inferred level of differentiation. The traditional histological diagnosis modes of brain tumors include: H&E staining; morphological phenotype under microscope; observation of ultra-subcellular structure under electron microscope; immunohistochemical staining analysis of characteristic protein expression, etc. However, there is prognostic heterogeneity among patients with the same grade and tissue type; patients with different grades and different tissue types have the same prognostic effect; all these indicate that the traditional histopathological diagnosis mode can no longer meet the requirements of brai...

AI Technical Summary

Problems solved by technology

[0006] First, the process of FISH detection of 1p / 19q chromosome deletion is cumbersome and takes a long time;

[0007] Second, the process of judging the results of FISH detection of 1p / 19q chromosome deletion is time-consuming and laborious, and each needs to calculate the number of fluorescence in 100 cells

[0008] Third, the diagnosis of glioma not only requires the detection of molecular markers such as 1p / 19q by FISH technology, but also the detection of molecular markers such as IDH, TERT, and EGFR by other detection technologies. Single-use detection of molecular markers

[0011] The second MLPA can only be used to detect gene deletion or duplication, such as 1p / 19q deletion, but it cannot meet the detection of other molecular markers of glioma, such as IDH, TERT and EGFR, etc.

Real-time fluorescence quantitative PCR microsatellite analysis technology detects 1p / 19q chromosomal deletion in glioma. The disadvantage is also that it cannot meet the one-time detection of other molecular markers of glioma, such as IDH, TERT and EGFR.

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