Construction method and applications of tumor BRCA1/2 gene variation library for high-throughput sequencing detection

Abstract

The present invention discloses a construction method of a tumor BRCA1 / 2 gene variation library for high-throughput sequencing detection, wherein the tumor BRCA1 / 2 gene variation library covers the mutations at all the exon regions of the human BRCA1 / 2 gene. According to the present invention, the complete exon sequences of the BRCA1 gene and the BRCA2 gene are subjected to single-tube amplification to rapidly complete the construction of the library, wherein the construction time of the whole library is only 2-3 h, and the manual time is only 15-30 min; and by combining the obtained library and the high-throughput sequencing platform, the difficult problem that the detection of the complete exon sequence is required on the basis of the small amount of the clinical samples in tumor diseases in the clinic can be effectively solved, and the cost is low.

Description

technical field [0001] The invention specifically relates to a method for constructing a tumor BRCA1 / 2 gene variation library for high-throughput sequencing detection and its application. Background technique [0002] Breast cancer and ovarian cancer are the most common malignant tumors that threaten women's health. The incidence of breast cancer in women varies significantly in different countries, with the highest incidence in the United States and Northern Europe. The occurrence and development of breast cancer are related to many factors, which are the result of the joint action of environmental factors and genetic factors. BRCA1 and BRCA2 are the abbreviations of Breast Cancer Susceptibility Gene1 and 2, that is, breast cancer susceptibility gene 1 / 2. Among them, the BRCA1 gene is located on human chromosome 17q21 and consists of 23 exons; the BRCA2 gene is located on human chromosome 13q12.3 and consists of 27 exons. Both BRCA1 and BRCA2 genes are tumor suppressor g...

AI Technical Summary

Problems solved by technology

[0006] Existing tumor gene detection mainly uses fluorescent quantitative PCR method, which has a low detection throughput. For the whole exome detection of BRCA1 and BRCA2 genes, the cost is high and the sample size is large, resulting in high clinical fees and insufficient sample size.

At present, the industry-recognized gold standard for gene mutation detection is the Sanger sequencing method. This type of technology is complex to operate, has low throughput, low sensitivity, and has a certain risk of missed detection. It is difficult to meet the detection needs of a large number of tumor patients.

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