A primer combination, method and kit for detecting large fragment recombination of brca1 and brca2 genes

Abstract

The invention discloses primer compositions, method and kit for detecting recombination of large fragments of BRCA1 and BRCA2 genes. Each pair of primers in the primer composition can obtain a characteristic fragment from the region, where at least one exon of the BRCA1 or BRCA2 gene is located, through amplification, and the length of the characteristic fragments vary; after optimization, the primers were divided into five groups for multiple PCR amplification, and the length of each pair of amplified products is different; multiplex PCRs are carried out in the form of a GeXP-PCR, linear specific amplification is conducted on the characteristic fragments in the early stage of a PCR, and indifference amplification is conducted on the characteristic fragments by universal primers in the late stage of the PCR; one of the universal primers can carry suitable fluorescence-modified groups for analysis on corresponding capillary electrophoresis analysis equipment so as to obtain relative content information of each characteristic fragment, and thus information of large fragment recombination mutation in a corresponding gene region is obtained. The method for detecting the recombination of the large fragments of the BRCA1 and BRCA2 genes has the advantages of being simple to operate and low in cost; compared with an ordinary end point multiplex PCR, the problem of multiple amplification preference is solved, and the reliability is improved.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a primer combination, a method and a kit for detecting the recombination of large fragments of BRCA1 and BRCA2 genes. Background technique [0002] BRCA1 and BRCA2 are important tumor suppressor genes involved in the process of chromosomal damage repair. Mutations in BRCA1 and BRCA2 may result in reduced or absent gene function, thereby increasing the risk of cells becoming cancerous. Existing data show that germline mutations in these two genes are the most important factors in causing hereditary breast and ovarian cancer, and mutation carriers have an 87% chance of developing breast cancer and a 44% chance of ovarian cancer in their lifetime. . Therefore, the detection of BRCA1 / 2 gene mutations in high-risk groups has important guiding significance for the prevention, discovery and treatment of cancer. [0003] There are two types of mutations in BRCA1 / 2. The first is t...

AI Technical Summary

Problems solved by technology

The first is the mutation of a single base or the deletion and insertion of a small number of bases, which can cause changes in the amino acid sequence, premature termination of translation, or abnormal processing of messenger RNA, thereby affecting the function of the protein

The second is large fragments of DNA recombination, which can cause deletion or duplication of one or more exons, thereby disrupting gene function

MLPA technology is widely used in the detection of BRCA1 / 2 and many other gene large fragments of DNA recombination. At present, there are still shortcomings of large data variation, and the detection accuracy needs to be improved.

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