Human BRCA1/2 gene variation detection quality control material, preparation method thereof and application of material

Abstract

The invention provides a preparation method of a human BRCA1/2 gene variation detection quality control material, and belongs to the technical field of high-throughput sequencing. After a plurality ofBRCA1/2 gene variation positive and stably transferred human tumor cell line genomes DNA (deoxyribonucleic acid) are diluted, variation sites on BRCA1/2 genes in diluents are measured by a high-throughput sequencing method, one of the variation sites is optionally selected as a positive control site, and a wild type site of the variation sites is selected as a negative control site. The copy number of the genomes in the diluents is quantified by a real-time fluorescent quantitative PCR (polymerase chain reaction) method. The mixing ratio of the human tumor cell line genome DNA diluents is calculated according to the copy number of the genomes in the diluents and the target allele frequency of the selected variation sites. The human tumor cell line genome DNA diluents are mixed according to the mixing ratio to obtain the human BRCA1/2 gene variation detection quality control material.

Description

technical field [0001] The invention belongs to the technical field of high-throughput sequencing, and in particular relates to a human BRCA 1 / 2 gene variation detection quality control product and its preparation method and application. Background technique [0002] Breast cancer is one of the most common malignant tumors that seriously affects women's health and even threatens their lives. According to statistics, the incidence of breast cancer accounts for 7-10% of the incidence of various malignant tumors in the whole body. Breast cancer can be divided into sporadic and hereditary two categories. Among them, hereditary breast cancer accounts for about 5-10% of the incidence of breast cancer. Breast cancer susceptibility genes BRCA1 (the first familial breast and ovarian cancer susceptibility gene) and BRCA2 (the second familial breast cancer susceptibility gene) have heritable mutations, and the risk of developing breast cancer in the lifetime of BRCA1 or BRCA2 mutatio...

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Problems solved by technology

Existing detection of fetal chromosomal aneuploidy (T21, T18, T13) uses a semiconductor sequencing method. Due to the different purposes of the analysis, such detection analyzes the variation at the chromosome level, so it cannot be used to target gene variation as Purpose Detection Analysis

Other gene mutation detection products, such as EGFR hotspot mutation detection, use PCR-fluorescent probe method, fluorescent PCR-capillary electrophoresis method, flow fluorescent hybridization method, FISH sequencing method, Taqman-ARMS method, etc., because they can only detect The detection efficiency is low and the cost is high for the mutation of a specific site, but there is no specific site for the mutation of the BRCA1 / 2 gene, and the incidence of the mutation has no obvious site specificity, so the conventional method is not suitable for this type of mutation However, it cannot meet the needs of BRCA1 / 2 gene detection, which requires detection and analysis of the entire coding region of the target gene.

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