Primer, method and kit for detecting mutation sites of human BRCA1 and BRCA2 gene all-coding sequences

Abstract

The invention discloses a primer, a method and a kit for detecting mutation sites of human BRCA1 and BRCA2 gene all-coding sequences. The primer comprises forward and reverse primers which can amplifyfor covering all the 24 coding zones of BRCA1 gene and of which base sequences are shown as SEQ ID NO:3-52. The primer further comprises forward and reverse primers which can amplify for covering allthe 27 exon coding zones of BRCA2 gene and of which base sequences are shown as SEQ ID NO: 53-134. A detection method based on NGS technique comprises the following steps: combining a specific primerwith a target template DNA sequence; adopting a universal primer for amplifying a to-be-detected sample target area; purifying a library by using a magnetic bead; performing high-throughput sequencing on the acquired library and analyzing mutation of BRCA1 and BRCA2 genes. The primer, method and kit disclosed by the invention can cover all the to-be-detected mutation sites; uniformity is high; experimental procedures can be simplified; detection sensitivity is promoted; a guidance is supplied for selective medication of patients suffering from familial breast cancer and ovarian cancer as wellas for genetic predisposition; morbidity risk is effectively reduced.

Description

technical field [0001] The invention relates to the field of gene mutation detection in biotechnology, and more specifically relates to a primer, method and kit for detecting mutation sites in the full coding sequences of human BRCA1 and BRCA2 genes. Background technique [0002] The high incidence of breast cancer has seriously threatened women's health and is one of the most common cancers in women. Breast cancer can be divided into two categories: sporadic and hereditary. At present, it is believed that most hereditary breast cancers are caused by gene mutations, among which BRCA1 and BRCA2 are the most direct breast cancer susceptibility genes discovered so far. Studies have shown that people with BRCA1 and BRCA2 gene mutations have a higher risk of developing breast cancer and recurrence than the general population. Women with BRCA1 and BRCA2 gene mutations have a lifetime risk of breast cancer as high as 87%. . Not only that, but the BRCA1 / 2 gene is also a susceptibi...

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a primer, method and kit for detecting mutation sites in the full coding sequence of BRCA1 and BRCA2 genes related to human genetic susceptibility based on multiplex PCR and high-throughput sequencing technology, aiming to solve the current There are problems such as high cost, many steps, low sensitivity, long cycle and easy pollution in the technology

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