A detection primer, kit and detection method for mitochondrial dna gene mutation of leber hereditary optic neuropathy

Abstract

The invention relates to a detection primer, a kit and a detection method for the mutation of the mitochondrial DNA gene of Leber's hereditary optic neuropathy. Based on the principle of Sanger sequencing, the present invention has the characteristics of high sensitivity, stability and accuracy, can simultaneously detect the four most common pathogenic mutations of LHON mtDNA, and solves the low sensitivity and the The problems of insufficient specificity, easy pollution and high cost provide LHON with an accurate genetic diagnosis method, which is of great significance for genetic counseling and gene therapy of the disease.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a detection primer, a kit and a detection method for the mutation of the mitochondrial DNA gene of Leber's hereditary optic neuropathy. Background technique: [0002] Leber's hereditary optic neuropathy (LHON) is one of the most common mitochondrial genetic diseases and one of the most common causes of vision loss in young adults [Man PYW et al. J MedGenet, 2002]. More than 95% of patients are caused by three primary pathogenic mutations in mitochondrial DNA (mtDNA), including m.11778G>A in the MT-ND4 gene, m.14484T>C in the MT-ND6 gene and MT-ND1 gene m.3460G>A [Cerelli V et al. Prog Retin Eye Res, 2004]. The m.3635G>A mutation located in the MT-ND1 gene is the fourth common mutation in the Chinese population, and its mutation frequency is comparable to the m.3460G>A mutation [Jia XY et al. J Hum Genet, 2006]. Mutation detection of these four...

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Problems solved by technology

[0003] At present, on the one hand, there is no commercial LHON common mtDNA 4-site mutation detection kit on the market; on the other hand, the most common method for the genetic diagnosis of LHON has allele-specific PCR (AS-PCR) [CN1288254C, CN102146443A , CN101781683A], restriction fragment length polymorphism (RFLP) [CN1143117A, CN102747152A, CN104774927A, CN103173545A, CN104774926A, CN103173544A] and allele-specific fluorescent probe technology PCR [CN1721839A, CN1712543A common, detect single or 3] Mutation sites, these detection methods have the advantages of simplicity, rapidity, and low cost. However, due to the high frequency of mtDNA mutations in the population, mutations located in the vicinity of the primary mutation will affect the results, resulting in false positives or false negatives. In addition, There are also chip detection methods [CN101497925A, CN104805210A], which can detect more LHON-related mtDNA variations at the same time, and have the advantages of accurate and high-throughput, but this method is expensive

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