Primers, kits and methods for detecting human brca1 and brca2 gene mutations

Abstract

The invention discloses primers, a kit and a method for detection of human BRCA1 and BRCA2 gene mutation. The upstream primers are in a sequence shown as SEQ ID NO: 1,3,5,7,9-193, and meanwhile the universal primer Tag1 is added to the 5' end of the sequence; the downstream primers are in a sequence shown as SEQ ID NO: 2,4,6,8,10-194, and meanwhile the universal primer Tag2 is added to the 5' end of the sequence. The upstream primers with a P5 sequence and a sequence shown as SEQ ID NO: 221-236 and the downstream primers with a P7 sequence and a sequence shown as SEQ ID NO: 197-220 are also included. Through the primers, the kit and the method, mutation of all exons of BRCA1 and BRCA2 genes, a connection region between the exons and introns, an untranslated region and a promoter region can be detected rapidly, accurately, easily and conveniently.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to primers, kits and methods for detecting human BRCA1 and BRCA2 gene mutations. Background technique [0002] Breast cancer and ovarian cancer are common malignant tumors in women. The lifetime risk of breast cancer and ovarian cancer in the general population is 13% and 1.5%, respectively. Increasing at a rate of more than 3% year by year (Antoniou A, Pharoah PD, Narod S, et al. Average risks of breast and ovarian cancer associated with BRCA1 or BRCA2 mutations detected in case series unselected for family history: A combined analysis of 22 studies. American Journal of Human Genetics 2003;72(5):1117–1130). At present, it is believed that most hereditary breast cancers are caused by gene mutations, and the mutations of BRCA1 and BRCA2 genes are closely related to the onset of breast cancer. Studies have shown that women carrying BRCA1 or BRCA2 gene pathogenic mutations have a 5-30-fo...

AI Technical Summary

Problems solved by technology

[0005] At present, there are many methods for detecting BRCA1 / 2 gene mutations, mainly including: fluorescent quantitative PCR technology, which has high sensitivity and strong specificity, but it can only detect one type of mutation each time, and cannot completely cover the entirety of BRCA1 and BRCA2 genes. Coding region; restriction small fragment length polymorphism analysis (RFLP method), used to detect genes with altered restriction sites, can directly determine the genotype, but cannot be used for gene detection without new restriction sites, and The experimental operation is cumbersome, the detection cycle is long, and the cost is high; the Sanger sequencing method, the sequence close to the primer is easy to be inaccurate, and the sequencing cycle is long, the operation is complicated, and the cost is expensive, which is difficult to meet the needs of practical applications

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