Construction method and application of gastrointestinal stromal tumor polygenic variation library

Abstract

The invention discloses a construction method and an application of gastrointestinal stromal tumor polygenic variation library. In the method, monotube is performed to a plurality of target sequences, so that target area capturing before the construction of the library is quickly completed, wherein the whole target area capturing process only needs 2-3 h, manual time being only 15-30 min. With combination of a high-throughput sequencing platform and other library construction universal kits, the method solves a difficult that detection on multiple genes and target points is required in clinical treatment of the gastrointestinal stromal tumor on the basis of a less quantity of clinical samples at present. The method is low in cost and can be applied in various high-throughput sequencing platforms, gene chip platforms and hybridization detection platforms in the prior art.

Description

technical field [0001] The present invention relates to a construction method and application of a multi-gene variation library for gastrointestinal stromal tumors, in particular to a construction method for high-throughput sequencing detection of a multi-gene variation library for gastrointestinal stromal tumors, a kit and a method thereof. application. Background technique [0002] Gastrointestinal stromal tumors (GIST) are a type of tumor originating from the mesenchymal tissue of the gastrointestinal tract, accounting for 1-3% of gastrointestinal malignant tumors, and the estimated annual incidence is about 10-20 / 100 Ten thousand, mostly in middle-aged and elderly patients, rare in patients under 40 years old, and there is no significant difference in incidence between men and women. [0003] Studies have found that the occurrence of GIST is mainly related to the continuous activation of tyrosine kinase caused by the mutation of c-KIT gene, which makes the proliferation...

AI Technical Summary

Problems solved by technology

At present, the detection of gene mutations mainly relies on the PCR-direct sequencing method, but the PCR-direct sequencing method can only amplify a pair of primers in one PCR reaction, which takes a long time and has low sequencing sensitivity, and can only detect mutations with a frequency of more than 10%.

At present, gastrointestinal stromal tumors are required to detect more genes and more targets in addition to detecting KIT and PDGFRA, involving 80 thermal mutation sites. Continue to use PCR-direct sequencing method, which is labor-intensive, cumbersome and sensitive The degree is low, and it cannot be completed in actual work.

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