NRAS gene mutation detection specificity primer and liquid chip thereof

Abstract

The invention discloses an NRAS gene mutation detection specificity primer and a liquid chip thereof. The liquid chip mainly comprises: an ASPE primer composed of a 5'-terminal tag sequence and 3'-terminal specificity primer sequences focused on target gene mutation sites, wherein the specificity primer sequences comprise SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20, SEQ ID NO.21 and/or SEQ ID NO.22 focused on a Codon12 site, SEQ ID NO.23, SEQ ID NO.24, SEQ ID NO.25, SEQ ID NO.26 and/or SEQ ID NO.27 focused on a Codon13 site, SEQ ID NO.28 and SEQ ID NO.29 focused on a Codon18 site, and/or SEQ ID NO.30, SEQ ID NO.31, SEQ ID NO.32, SEQ ID NO.33 and/or SEQ ID NO.34 focused on a Codon61 site; a microsphere coated by an anti-tag sequence; and an amplimer. The consistency between the detection result of the detection liquid chip provided by the invention and the detection result of a sequencing method is high to 100%, and the wild-type and mutant parallel detection 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 NRAS gene mutation detection and a liquid phase chip. Background technique [0002] The NRAS gene is a member of the RAS oncogene family and is located on chromosome 1. The total length of the gene is 85kb, and its cDNA is about 2.1kb long. It encodes a protein containing 495 amino acids, called p21Ras. RAS proteins have GTP / GDP binding ability and GTPase activity, and their normal function is G protein-like regulatory proteins, which have signal transduction from membrane-bound receptors to adenylate cyclization process, and participate in the normal regulation of cell cycle . Current studies have shown that N-ras gene mutations have the highest frequency among ras gene mutations, and mutations at the 12th / 13th or 61st codons are the most common. Ras gene mutations are closely related to the prognosis of MDS. [0003...

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.

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

Images