High throughput sequencing method for detecting genes of multiple mutant types and applications thereof

Abstract

The invention discloses a high throughput sequencing method for detecting genes of multiple mutant types and applications thereof, and belongs to the field of gene detection. By searching database, variation sites of genes for a cancer target treatment, chemotherapy drug resistant genes, and chemotherapy drug sensitive genes, 100 bp of downstream of the variation sites, and 100 bp of upstream of the variation sites are taken as a target section; a capture probe matched with the target section is synthesized, the target area of DNA of a sample is captured, a high throughput sequencing library is established; then high throughput sequencing is carried out to obtain the sequence of a target section; and the results of cancer related gene mutation, insertion mutation, delete mutation, mutationof copy number, and gene fusion mutation can be obtained. The provided method can detect multiple kinds of mutations of 105 cancer related genes at a time and has the characteristics of high capturing efficiency and good uniformity.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and more specifically relates to a high-throughput sequencing method for detecting multiple mutation types of genes and its application, which can simultaneously detect multiple cancer-related gene fusions, copy number variations, point mutations and insertions Deletion variant. Background technique [0002] In China, the death rate caused by tumors is increasing year by year. Cancer has become an important factor affecting public health. In 2015, statistics showed that there were 4,292,000 new cancer patients and 2,814,000 deaths. With the popularization of diagnosis, the proportion of lung cancer diagnosis and death rate is increasing. Cancer rates are higher in rural areas than in urban areas. The tumor incidence rate in the United States is 45%, and the mortality rate is 30%, and this ratio is also increasing year by year. However, with the application of early tumor screening, di...

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Problems solved by technology

[0012] In summary, most of the gene detection methods in the prior art focus on the detection of point mutations, insertions / deletions, DNA copy number changes and other mutation types, while it is rare to combine multi-oncogene fusion problems with point mutations, insertions / deletions, DNA copy number changes, etc. There are reports of simultaneous detection of multiple changes, the limitation of clinical detection range (mutation detection of several sites) and the high detection cost of Sanger sequencing technology for multi-gene detection, which greatly limits its practical clinical application.

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