Specific primers and liquid-phase chip for detection of brca1 gene mutation

Abstract

The invention discloses a BRCA1 gene mutation detection liquid chip, and specific primers. The BRCA1 gene mutation detection liquid chip mainly comprises: ASPE primers, wherein each ASPE primer is composed of 5'-terminal tag sequence, and 3'-terminal specific primer sequence targeting target gene mutation sites, and the specific primer sequence comprises SEQ ID No.13 and SEQ ID No.14 targeting A926G site, SEQ ID No.15 and SEQ ID No.16 targeting C2077T site, SEQ ID No.17 and SEQ ID No.18 targeting A1525G site, SEQ ID No.19 and SEQ ID No.20 targeting G53675A site, SEQ ID No.21 and SEQ ID No.22 targeting C36048T site, and / or SEQ ID No.23 and SEQ ID No.24 targeting C2612T site; microballoons coated with different anti-tag sequences; and amplification primers. Self-agreement ratio of detection results of the liquid chip with detection results of sequencing is as high as 100%; and parallel detection of wild types and mutant types of a plurality of mutation sites is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for BRCA1 gene mutation detection and a liquid phase chip. Background technique [0002] The BRCA1 gene is called breast cancer 1 (breastcancer1, earlyonset, BRCA1), located on the long arm of chromosome 17 17q21, about 100 kb, containing up to 41.5% of Alu repeats and 4.8% of other repeats. Contains 24 exons, 22 of which are transcribed into 716kb mRNA, which can finally encode a protein with 1863 amino acids. The 11th exon is larger, 314kb long, accounting for 61% of the entire coding region. The N-terminal sequence of the BRCA1-encoded protein contains a ring domain, which can form a ring-2 ring heterodimer with BRCA12 associated ring domain protein (BARD1). It is generally believed that the N-terminal of BRCA1 regulates the function of RNA synthetase plays an important role. The BRCA1 gene is a member of the tumor su...

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of BRCA1 gene mutation mainly include: fluorescent quantitative PCR technology, PCR-RFLP technology, direct sequencing method. The disadvantage of high positive rate, and only one mutation type can be detected at a time

The PCR-RFLP method is based on the change of the restriction endonuclease recognition site caused by gene mutation, such as the loss of the site or the generation of a new site, a specific fragment is amplified by PCR, and then cut and amplified with a restriction endonuclease. Amplify the product and observe the size of the fragment by electrophoresis. This method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype. However, this method cannot be used for the detection of gene mutations without new restriction sites.

However, the sequence near the primers at both ends of the direct sequencing method is easy to be inaccurate, and the sequencing method is costly and complicated to operate. For those samples with a mutation ratio of less than 15% to 20%, it is difficult to find mutations if the sequencing method is used for detection. Therefore, these detection methods are difficult to meet the needs of practical applications.

Images