Quick detection method for IDH2 gene mutation

Abstract

The invention provides a quick detection method for IDH2 (isocitrate dehydrogenase) gene mutation, which comprises the following steps that a pair of wild type primers is designed at the two ends of an IDH2 gene mutational site; a pair of mutational type primers is designed at the mutational site; the wild type primers and the mutational type primers of an IDH2 gene are designed respectively; the primers are amplified to be a corresponding wild type template and a mutational type template respectively; the two templates are subjected to PCR (polymerase chain reaction) amplification by the wild type primers to be a corresponding wild type target fragment and a mutational type target fragment respectively; the two fragments are mixed in different proportions; and an HRM (high resolution melting) method is adopted to for distinguishing. Compared with other methods, the quick detection method has the advantages of high specificity, high sensitivity, convenience and quickness, and is higher in flux and lower in single cost.

Description

Technical field: [0001] The invention relates to the fields of biotechnology and medicine, especially molecular biology, molecular diagnosis and real-time quantitative PCR technology. Background technique: [0002] Isocitrate Dehydrogenase (IDH) converts isocitrate into α-ketoglutarate in the tricarboxylic acid cycle, which plays an important role in the body's energy metabolism, biosynthesis and anti-oxidative stress . There are two types of IDH enzymes in the human body: IDH enzymes that require NAD as a coenzyme (EC1.1.1.41), and IDH enzymes that require NADP (EC1.1.1.42); the two are in different subcellular Parts catalyze the same reaction: NADP+ is used as coenzyme in cytosol, and NAD+ is used as coenzyme in inner mitochondrial membrane. [0003] Determination of serum IDH enzyme content has certain significance in clinical diagnosis of liver disease, especially in patients with malignant tumors, elevated serum IDH enzyme is often a signal of liver metastasis. In ad...

AI Technical Summary

Problems solved by technology

[0015] First: the sensitivity is not high, and a low proportion of mutations (<20%) cannot be detected;

[0016] Second: it takes a long time, usually 2-3 days;

[0017] Third: high cost

Images