A hybrid capture kit and method for detection of mutations in breast cancer susceptibility genes brca1 and brca2

Abstract

The invention discloses a hybrid capture kit for detecting mutation of breast cancer susceptibility genes BRCA1 and BRCA2. The hybrid capture kit comprises a hybridization reagent, a PCR amplification reagent and an enrichment degree detection reagent, wherein the hybridization reagent comprises probe mixtures from SEQ NO.1 to SEQ NO. 173, and the mole ratio is SEQ NO.1: SEQ NO. (2-172): SEQ NO. 173=1: 1: 1. During detection, the use amount of each of the probe mixtures with the concentration of 2nM is 1 [mu]l. The kit disclosed by the invention can be used for high-sensitivity, high-flux, low-cost and easy-operation full-explicit mutation detection for the genes BRCA1 and BRCA2, and can assist a clinical doctor in screening out individuals with the breast cancer susceptibility risk to fulfill the aim of preventing occurrence of breast cancers.

Description

technical field [0001] The invention relates to the field of gene mutation detection, in particular to a hybrid capture kit and a detection method for detection of breast cancer susceptibility genes BRCA1 and BRCA2 mutations. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, known as the female killer, and its incidence rate is second only to lung cancer. Among them, hereditary breast cancer accounts for about 5% to 7% of the incidence of breast cancer. The greatest risk factor for breast and ovarian cancer is an inherited mutation in the breast cancer susceptibility gene BRCA1 or BRCA2. BRCA1 and BRCA2 are two tumor suppressor genes. Abnormalities in their structure and function are closely related to the occurrence of breast cancer and ovarian cancer. They can not only inhibit cell growth, but also participate in cell cycle regulation, gene transcription regulation, DNA damage repair and apoptosis, etc. A variety of importan...

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that a specific variant cannot be judged through the melting curve graph, and the detection in the downstream analysis still needs to be supplemented by sequencing, etc.

[0005] Multiplex PCR amplification capture technology uses multiplex PCR to amplify and enrich the DNA fragments in the target region of the sample genome, and then perform library construction and high-throughput sequencing. The disadvantage of this technology: the throughput is small, and the longest genomic DNA can be obtained by one PCR at present. The fragment length should not exceed 50kb, and requires special enzymes and special PCR conditions, high cost and poor stability

This method has the following disadvantages: RFLP experimental operation is cumbersome, the detection cycle is long, the cost is high, there are false positives caused by incomplete digestion in the first round, non-closed tube operation, easy to contamination, and difficult to meet the clinical detection requirements

[0007] The direct DNA sequencing method has the following disadvantages: long detection period, high cost, non-closed tube operation, it is difficult to avoid cross-contamination, and the throughput is not high

In addition, the sensitivity of this method is low, and problems such as heterozygous mutations, gel compression, and the existence of GC-rich regions make it difficult to obtain accurate data through one-time sequencing, and repeated sequencing is required to avoid false positives. Therefore, the direct sequencing method Difficult to apply to clinical testing

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