Specific primers and liquid chip kit for xrcc1 gene mutation detection

Abstract

The invention discloses an XRCC1 (X-Ray Repair Cross-Complementing Gene 1) gene mutation detection liquid-phase chip and specific primer. The XRCC1 gene mutation detection liquid-phase chip mainly comprises an ASPE primer, microspheres and an amplification primer, wherein the ASPE primer consists of a tag sequence at a 5'-end and a specific primer sequence for a gene mutation site at a 3'-end; the specific primer sequence is SEQ ID NO. 7 and SEQ ID NO. 8 for an A1196G site, SEQ ID NO. 9 and SEQ ID NO. 10 for a C580T site, and SEQ ID NO. 11 and SEQ ID NO. 12 for a G839A site; the microspheres are coated by an anti-tag sequence. The agreement rate of the detection result of the XRCC1 gene mutation detection liquid-phase chip and that of a sequencing method is as high as 100%. Independent or parallel detection on a wild type and a mutation type of multiple mutation sites can be achieved.

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 detecting XRCC1 gene mutation and a liquid phase chip kit. Background technique [0002] Human X-ray repair cross-complementing gene 1 (XRCC1) is the first isolated mammalian gene that affects the sensitivity of cells to ionizing radiation, and it is widely involved in the repair of DNA damage. [0003] XRCC1 is located on human chromosome 19q13.2-19q13.3, with a size of 33kb and 17 exons. The purified XRCC1 protein consists of 633 amino acid residues. As a scaffolding protein, XRCC1 directly forms a complex with polymerase β, DNA ligase III and poly ADP ribose polymerase [poly(ADP ribose) polymerase, PARP], and jointly participates in the base excision caused by ionizing radiation and oxidative damage repair and single-strand break repair. Many studies have shown that individual XRCC1 polymorphic sites have mutation...

AI Technical Summary

Problems solved by technology

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.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry is a soft ionization technology that has powerful and mature functions in the detection of protein and other biological macromolecules. However, in the field of nucleic acid detection, due to the particularity of nucleic acid molecules, detection is subject to certain limit

Although immunohistochemistry has the advantages of specificity, strong sensitivity, and simple operation, there is no uniform standard for the use of antibodies, operating procedures, and result judgments, which affects its clinical promotion and application.

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