Multiplex-PCR (polymerase chain reaction) detection method and kit for BRCA1 and BRCA2 gene mutation

Abstract

The invention provides a method and a kit for detecting BRCA1 and BRCA2 gene mutation. The method and the kit utilize two sets of PCR (polymerase chain reaction) primers, wherein a first set of PCR primers comprises 189 pairs of PCR primers and is divided into two groups, a first group comprises 95 pairs of primers with sequences represented as SEQ ID NO.5-SEQ ID NO.194, a second group comprises 94 pairs of primers with sequences represented as SEQ ID NO.195-SEQ ID NO.382, a sequence represented as SEQ ID NO.1 is added to the 5' end of each upstream primer, and a sequence represented as SEQ ID NO.2 is added to the 5' end of each downstream primer; sequences of upstream and downstream primers of a second set of PCR primers are represented as SEQ ID NO.3 and SEQ ID NO.4.

Description

technical field [0001] The invention relates to the field of gene mutation detection, in particular to multiplex PCR detection of gene mutation. Background technique [0002] BRCA1 and BRCA2 are two genes that can inhibit the occurrence of malignant tumors, and play an important role in regulating the replication of human cells, the repair of DNA damage in genetic material, and the normal growth of cells. Hundreds of BRCA1 and BRCA2 mutations have been identified, some of which are associated with hereditary breast and ovarian cancers. Summarizing the lifetime risk of cancers associated with BRCA1 and BRCA2 gene mutations shows that those with BRCA1 gene mutations have a risk of breast cancer and ovarian cancer of 50%-85% and 15%-45%, respectively, and those with BRCA2 gene mutations have breast and ovarian cancer risks. The risk of ovarian cancer is 50%-85% and 10%-20%, respectively. These are high cancer odds compared to the average woman. [0003] Since the mutations o...

AI Technical Summary

Problems solved by technology

The RFLP method is a method that combines PCR with restriction enzyme digestion. This method has the following disadvantages: cumbersome experimental operations, long detection cycle, high cost, false positives caused by incomplete first round of enzyme digestion, non-closed tube operation, Easy to pollute and difficult to meet clinical testing requirements

The direct DNA sequencing method has the following disadvantages: long detection cycle, high cost, non-closed tube operation, difficulty in avoiding cross-contamination, and low throughput

However, the multiplex PCR used in this patent is only limited to some mutation sites on some exons of BRCA1 and BRCA2 genes, which obviously cannot cover hundreds of mutations in multiple exons of BRCA1 and BRCA2 genes. Therefore, this patent has relatively large limitations in use