Mitochondrial diseases MELAS and MERRF syndrome related mtDNA mutation site detecting gene chip and detecting method

Abstract

The invention relates to a mutation site examination of mitochondrial diseases MELAS and MERRF syndrome correlated mtDNA, disclosing a mutation site detection gene chip correlated with the syndrome mtDNA of the mitochondrial diseases MELAS and MERRF, wherein, a specific oligonucleotide probe is fixed on the surface lattice of the gene chip carrier and is used for detecting the mtDNA mutation sites correlated with mitochondrial diseases syndrome MELAS and MERRF. The invention also further discloses a preparation and application method of the gene chip. The gene chip of the invention can be used for detecting all mutation sites mtDNA concerned with the mitochondrial disease syndrome MELAS and MERRF with high sensibility and specificity and good repeatability; compared with the traditional molecular biological methods, the testing time is greatly shortened, and the purposes of clinical diagnosis and identification diagnosis of the MELAS and MERRF syndrome are realized.

Description

technical field [0001] The invention relates to the detection of mtDNA mutation sites related to mitochondrial disease MELAS and MERRF syndrome, in particular to a gene chip for detecting mtDNA mutation sites related to mitochondrial disease MELAS and MERRF syndrome and its production method, and using the chip to detect clinical samples of patients Detection and screening of mtDNA mutation sites associated with MELAS and MERRF syndromes in China. Background technique [0002] It is now known that mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) and myoclonic epilepsy with ragged red fibers (MERRF) ) are two common mitochondrial diseases mainly caused by mutations in mtDNA. MELAS is a group of mtDNA disorders with high clinical variability and genetic heterogeneity, most MELAS cases are caused by mt-tRNA Leu(UUR) Genetic heterogeneity mutation A3243G caused; a few are caused by other mtDNA point mutations (such as T3271C) or large fragm...

AI Technical Summary

Problems solved by technology

Although many mtDNA mutant detection methods have been established, such as restriction fragment length polymorphism (RFLP), single-strand conformation polymorphism (SSCP), specific primer sequence analysis (SSP), and conformation-sensitive electrophoresis (CSGE). , Transient Temperature Gradient Electrophoresis (TTGE), Denaturing Gradient Gel Electrophoresis (DGGE) and Denaturing High Performance Liquid Chromatography (DHPLC), etc., but there are still many limitations in practical application, such as high cost of RFLP, heavy workload and some important The mutation site cannot be detected because it is not recognized by the restriction endonuclease used, and other methods are more complicated than RFLP

More importantly, most of these methods can only detect one or a few mutation sites, and cannot simultaneously screen mutants with different genes at high throughput. The results of some methods still need to be verified by molecular sequencing

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