Breast cancer susceptibility gene BRCA1 and BRCA2 detection kit and method

Abstract

The invention discloses a breast cancer susceptibility gene BRCA1 and BRCA2 detection kit and method. The kit includes a first-round PCR reagent and a second-round PCR reagent. The first-round PCR reagent contains a buffer solution, magnesium salt, deoxyribonucleoside triphosphate, enzyme mixture liquid and a first-round PCR primer. The second-round PCR reagent contains a buffer solution, magnesium salt, deoxyribonucleoside triphosphate, enzyme mixture liquid and a second-round PCR primer. The first-round PCR primer includes 46 pairs of primers SEQ1-SEQ92 and 67 pairs of primers SEQ93-SEQ226, the 5'end of each upstream primer is added with a sequence SEQ227, and the 5'end of each upstream primer is added with a sequence SEQ228. The primer combination includes 113 pairs of primers and covers all the coding sequences and adjacent at least +5bp intron subdomains of a breast cancer susceptibility genes. An NGS technology is adopted to detect mutations of all the coding sequences and splice sites of the breast cancer susceptibility genes BRCA1 and BRCA2, the kit has very high specificity and accuracy, and the method is simple, convenient and efficient.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection kit and method for breast cancer susceptibility genes BRCA1 and BRCA2. Background technique [0002] Breast cancer and ovarian cancer are the most common malignant tumors that threaten women's health. BRCA1 and BRCA2 are two susceptibility genes that are closely related to breast cancer and ovarian cancer, and play important roles in DNA damage repair, cell cycle regulation, gene transcription activation, and chromatin stability. Studies have shown that about 2%-6% of female breast cancer patients and 10%-15% of ovarian cancer patients have BRCA1 and BRCA2 gene mutations. The mutation status of these two genes, BRCA1 and BRCA2, is closely related to the incidence of familial breast cancer and ovarian cancer. Individuals with BRCA1 and BRCA2 gene mutations have a significantly increased risk of developing breast and ovarian cancer and a significantly inc...

AI Technical Summary

Problems solved by technology

In terms of detection sensitivity, Sanger sequencing can generally only detect mutations with a mutation rate of more than 20%. For the detection of all coding sequences of BRCA1 and BRCA2 and adjacent intron sequences, the detection throughput is low, the cycle is long, and the cost of single-sample detection also higher

In addition, there are also detection kits based on High Resolution Melting Analysis (HRM) technology on the market, but they can only detect a few known sites, and their effect is very limited

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