Method and primers for detecting third exon mutation site of RUNX1 gene

Abstract

The invention discloses a method and primers for detecting a third exon mutation site of a RUNX1 gene. Both the primers and the method comprise a forward primer, a reverse primer and a pair of sequencing primers. By adopting a Sanger sequencing method and by utilizing the pair of sequencing primers, the method and the primer can be used for rapidly detecting mutation conditions of the third exon mutation site of the RUNX1 gene in an acute myeloid leukemia patient body.

Description

technical field [0001] The invention belongs to the field of life science and biotechnology, in particular to a primer and a method for detecting the third exon mutation site of RUNX1 gene. Background technique [0002] Acute myeloid leukemia (AML) is a malignant disease caused by cumulative acquired gene changes in myeloid hematopoietic stem / progenitor cells, resulting in changes in cell proliferation, differentiation and apoptosis pathways. [0003] RUNX1, also known as AML1, is a member of the RUNX transcription factor protein family and is the most common target site of leukemia chromosomal translocation. RUNX1 is a very important transcription factor that can bidirectionally (promote or inhibit) regulate hematopoietic-related factors, thereby regulating the differentiation, apoptosis and self-renewal of hematopoietic stem cells. RUNX1 point mutations can often occur in myelodysplastic syndrome (MDS), AML, chronic myelomonocytic leukemia (CMML), and less commonly in mye...

AI Technical Summary

Problems solved by technology

ssCp technology is relatively simple, but the restriction site is easily affected by gene variation, which affects the judgment of the result. Due to the technical limitations of the method itself, the detection sensitivity is low

Fluorescence quantitative PCR method was used to detect the mutation of exon 3 of RUNX1 gene. Since the detected exon sequence was long and involved multiple mutation sites, it was necessary to design multiple pairs of primers and probes. Three mutant probes were designed. Specifically bind to the 3 mutation sites covered by the third exon of the RUNX1 gene, and design 3 corresponding wild-type probes to bind to the unmutated DNA of the 3 sites respectively. To detect a DNA sample, 3 Tube PCR reaction, each tube reaction system includes 1 pair of amplification primers, 1 wild-type probe, 1 corresponding mutant probe, the amount of sample DNA is relatively large, and the detection cost is also high, which brings great advantages to clinical applications. Inconvenience

In addition, the existing technology only detects a single point mutation with a high probability of mutation, but does not detect the overall mutation of the third exon

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