Primer combination and its application for detecting pdgfra gene mutation in trace tissue

Abstract

The invention discloses a primer combination for detecting gene PDGFRA mutation in microtissue. The primer combination comprises a pre-amplification primer group, a primer group for detecting mutation of a twelfth exon, a primer group for detecting mutation of a fourteenth exon and a primer group for detecting mutation of an eighteenth exon. According to the method, it aims at obtaining a high-concentration DNA template through a trace of DNA by means of pre-amplification, and the mutation situation of a gene PDGFRA in a sample is detected through a direct sequencing method. The primer combination is applied to detection reagents for detecting mutation of the gene PDGFRA, can detect samples with the DNA concentration lower to 100 pg and can simultaneously detect all mutation of the twelfth exon, the fourteenth exon and the eighteenth exon of the gene PDGFRA. The detection method is high in sensitivity and specificity, low in cost and suitable for detection of mutation of the gene PDGFRA of a clinical cancer patient and has very high clinical application value.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a PDGFRA Primer combination for gene mutation detection and its application in tumor drug selection and disease diagnosis. Background technique [0002] PDGFR is a single-chain membrane glycoprotein with a molecular weight of 180kD. The extracellular ligand-binding region contains 5 immunoglobulin-like domains, with conserved cysteine ​​residues and a single transmembrane segment; an intracellular tyrosine kinase The break in the region is a hydrophilic amino acid insertion sequence. The receptor molecule is composed of two subunits, α and β. The mature PDGFR is in dimer steady state form (αα, αβ, ββ) and the corresponding isomer of ligand PDGF (platelet-derived growth factor, PDGF) (PDGF - AA, AB, BB) binding. Both PDGFR α and PDGFR β are expressed in colorectal cancer tissues. PDGFR α is distributed in normal colorectal tissues, polyps and tumor tissues; PDGFR β...

AI Technical Summary

Problems solved by technology

However, the application of the last base mismatch in ARMS technology cannot completely block the amplification of wild-type DNA, there is a risk of false positives, and it can only be detected for specific sites

In addition, such as high performance liquid chromatography, capillary electrophoresis, etc., require special equipment, and the operation is complicated, which is not conducive to large-scale clinical promotion

Nucleic acid sequence amplification (NASBA), self-sequencing sequence replication (3SR) and strand displacement replication (SDA) are all isothermal amplification methods, but their specificity for the amplification of target sequences is not strong

[0006] despite this, PDGFRA The detection method of mutation is still based on the detection of tumor tissue samples as the gold standard, but in clinical practice, the detection is often limited due to insufficient tissue samples

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