ERCC1 gene mutation detection specific primer and liquid phase chip kit

Abstract

The invention discloses an ERCC1 gene mutation detection liquid phase chip and specific primers. The liquid phase chip mainly comprises ASPE primers, microspheres and amplimers, wherein each kind of ASPE primers is formed by 5'end tag sequences and 3'end specific primer sequences aiming at gene mutation sites, the specific primer sequences are SEQ NO.11 and SEQ NO.12 aiming at T354C sites, SEQ NO.13 and SEQ NO.14 aiming at G262T sites, SEQ NO.15 and SEQ NO.16 aiming at G197T sites, SEQ NO.17 and SEQ NO.18 aiming at T931G sites and SEQ NO.19 and SEQ NO.20 aiming at A1187G sites; microspheres are wrapped with anti-tag sequences. The coincident rate of the detection result of the liquid phase chip and the sequencing method can be as high as 100%, and wide type and mutant type single and parallel detection of multiple mutation sites can be 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 detecting ERCC1 gene mutation and a liquid phase chip kit. Background technique [0002] Nucleotide excision repair cross complementing group 1 (Excision Repair Cross Complementing group 1, ERCC1) is located on human chromosome 19q13.2, with a length of 15 kb, encoding a protein consisting of 297 amino acids, and is an important member of the excision repair family. A key member of the NER system, involved in DNA strand cleavage and damage recognition. At the same time, ERCC1 gene polymorphisms are not only related to the efficacy of tumor chemotherapy, but also have an impact on prognosis. In the process of tumor treatment, the enhancement of DNA repair ability affects the efficacy of antitumor drugs and is prone to drug resistance; the decline of DNA repair ability helps the persistence of the function of platinum-DN...

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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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