Primer composition for detecting human BRCA1 and BRCA2 gene mutation based on NGS method and reagent of primer composition

Abstract

The invention discloses a primer composition for detecting human BRCA1 and BRCA2 gene mutation based on an NGS method. The primer composition comprises 263 pairs of first group of primers and 20 pairs of second group of primers, each pair of the first group of primers or the second group of primers comprises a forward primer sequence and a reverse primer sequence, each of the forward primer sequence and the reverse primer sequence of each first group of primers comprises a sequence capable of specifically amplifying an exon, an intron or an UTR region of a BRCA1 / BRCA2 gene, and both the forward primer sequence and the reverse primer sequence of each second group of primers comprise specific reference sequences capable of specifically amplifying human genomes. According to the invention, the pathogenic point mutation, the short deletion / insertion and the copy number variation of the BRCA1 / BRCA2 gene can be effectively detected, genome DNA samples can be detected, tumor FFPE samples can be detected, the purposes of the genetic risk evaluation of the ovarian cancer and the breast cancer, the PARPi inhibitor medication guidance, the prognosis evaluation and the like can be met, the whole process is simple and efficient, and the good prospect is provided.

Description

technical field [0001] The invention relates to the technical field of genetic compositions, in particular to a primer composition and a reagent thereof for detecting human BRCA1 and BRCA2 gene mutations based on an NGS method. Background technique [0002] According to the 2018 Global Cancer Statistics Report, new cases of female breast cancer worldwide accounted for 11.6% of the total, ranking first with lung cancer. Breast cancer is the most common cancer in women (incidence rate is 24.2%), the mortality rate of breast cancer is 15.0%, and the incidence rate and mortality rate both rank first in women's morbidity and death. Ovarian cancer has an incidence rate of 3.4% and a mortality rate of 4.4%, ranking eighth among women in terms of incidence and mortality. [0003] Studies have shown that about 15% of breast and ovarian cancer patients carry germline (germline, that is, inherited from parents' germ cells) BRCA1 and / or BRCA2 gene mutations. Women carrying such germli...

AI Technical Summary

Problems solved by technology

[0007] However, the products currently on the market cannot well meet the ever-increasing need for BRCA gene mutation detection

First of all, the next-generation sequencing technology can be divided into two categories: probe hybridization capture method and multiple amplification method according to the different library construction methods: the probe hybridization capture method is expensive and the operation is relatively complicated; Overlapping amplified fragments, and these overlapping fragments will interfere with each other, so they must be divided into 2 tubes or even 6-8 tubes for operation; no matter which route the product operation is time-consuming and laborious

Secondly, the specificity and sensitivity of existing products are generally insufficient, and the required sample input volume is high, usually hundreds of nanograms or even several micrograms of samples are required, which is unacceptable for tissue samples; Its original design problem often fails to detect somatic mutations in tumor tissues, especially in paraffin-embedded tissues (Formalin-Fixed and Parrffin-Embedded, FFPE); Germline or somatic mutation, which belongs to the "copy number variation" (copynumber variation, CNV) of the entire gene or some exons, conventional products cannot be effectively detected

Images