Drop-off ddPCR method and kit for quantitatively detecting NPM1 gene mutation

Abstract

The invention relates to a detection method and a kit for detecting NPM1 gene mutation based on drop-offddPCR; primers and probes thereof are designed according to an NPM1 gene DNA sequence; two wild probes are designed aiming at a 12th exon mutation hot spot of an NPM1 gene; one wild probe is located on the mutation hot spot; the other wild probe is located outside the mutation hot spot; when mutation such as insertion, replacement and deletion exists in the mutation hot spot, the wild probe located at the mutation hot spot cannot be tightly combined with a template, so that the kit can detect various mutations of the mutation hot spot of the 12th exon of the NPM1 gene only by using two pairs of primer probes; and the kit is high in sensitivity and can be used for MRD monitoring.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method and kit for quantitatively detecting NPM1 gene mutation. Background technique

[0002] Nucleophosmin 1 (nucleophosmin, NPM1) gene is located on human chromosome 5q35, with 12 exons in total, and its protein consists of 294 nucleotides. NPM1 mutation is one of the most common genetic mutations in acute myeloid leukemia (AML). NPM1 mutations are detected in 25%–35% of AML cases, over 50% in cytogenetically normal AML (CN-AML), and in myeloproliferative and myeloid dysplastic disorders There are also discoveries. So far, about 50 NPM1 exon 12 mutations have been found, most of which are insertions between nucleotides 863 and 864. NPM1 mutations are very stable during the disease process and thus are one of the important molecular markers for better diagnosis and disease monitoring in AML.

[0003] Minimal residual disease (MRD) refers to submicroscopic lesions that still ex...

Images