Leber遗传性视神经病相关的三种原发性线粒体突变的无创检测方法建立

Abstract
Figure/Table
References (0)
Related Citation (15)

Download: PDF (1473 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Objective: To establish a non-invasive detection assay for three Leber’s hereditary optic neuropathy (LHON) families with m.11778G>A, m.14484T>C and m.3460G>A mutations. Methods: Three patients (WZ1481, WZ1478, WZ1435) with m.11778G>A, m.14484T>C and m.3460G>A mutations who visited School of Ophthalmology and Optometry, Wenzhou Medical University were included in this study. Purity and concentration of genes from buccal cell samples via manual sampling and reagent kits were determined. The whole genomic DNA of samples carrying m.11778G>A, m.14484T>C and m.3460G>A mutation with LHON as well as wild type were used as templates, amplification of DNA fragments each containing the above three mutations by PCR. The PCR products were analyzed by DNA sequencing, Dot blot, and Southern blot. Results: There were no significant differences in DNA concentration between buccal cells and blood (P>0.05). However, the purity of buccal cell DNA extracted by manual sampling was lower than the buccal cell DNA and blood DNA extracted by reagent kits (P<0.05). The DNA yielding of buccal cell DNA was similar to that of blood DNA. The results of DNA sequencing, Dot blot and Southern blot were also highly consistent. Both groups were positive in the control group and negative in the mutant group. Conclusion: The results of DNA sequencing, Dot blot and Southern blot demonstrate that buccal cell DNA can be used for the screening and identification of three primary mitochondrial mutations of LHON. This study has established a convenient and non-invasive detection assay suitable for clinical determination in three primary mitochondrial mutations associated with LHON.

Key words： mtDNA Leber’s hereditary optic neuropathy mutation non-invasive buccal swab

Received: 10 January 2019

	Service
	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	WANG Min
	LYU Yuanyuan
	XUE Ling
	ZHENG Binjiao
	GUAN Minxin.

URL:

https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2019/V49/I6/412

[1] LEBER T. Ueber hereditaere und congenital angelegte seh-nervenleiden[J]. Graefes Arch Opthal, 1871, 17: 249-291.
[2] NEWMAN N J, LOTT M T, WALLACE D C. The clinical characteristics of pedigrees of Leber's hereditary optic neuropathy with the 11778 mutation[J]. Am J Ophthalmol, 1991, 111(6): 750-762.
[3] WALLACE D C, SINGH G, LOTT M T, et al. Mitochondrial DNA mutation associated with Leber's hereditary optic neuropathy[J]. Science, 1988, 242(4884): 1427-1430.
[4] HOWELL N, OOSTRA R J, BOLHUIS P A, et al. Sequence analysis of the mitochondrial genomes from Dutch pedigrees with Leber hereditary optic neuropathy[J]. Am J Hum Genet, 2003, 72(6): 1460-1469.
[5] YU-WAI-MAN P, GRIFFITHS P G, HUDSON G, et al. Inherited mitochondrial optic neuropathies[J]. J Med Genet, 2009, 46(3): 145-158.
[6] MACKEY D, HOWELL N. A variant of Leber hereditary optic neuropathy characterized by recovery of vision and by an unusual mitochondrial genetic etiology[J]. Am J Hum Genet, 1992, 51(6): 1218-1228.
[7] HUOPONEN K, VILKKI J, AULA P, et al. A new mtDNA mutation associated with Leber hereditary optic neuroretinopathy[J]. Am J Hum Genet, 1991, 48(6): 1147-1153.
[8] HOWELL N, BINDOFF L A, MCCULLOUGH D A, et al. Leber hereditary optic neuropathy: identification of the same mitochondrial ND1 mutation in six pedigrees[J]. Am J Hum Genet, 1991, 49(5): 939-950.
[9] JOHNS D R, NEUFELD M J, PARK R D. An ND-6 mitochondrial DNA mutation associated with Leber hereditary optic neuropathy[J]. Biochem Biophys Res Commun, 1992, 187(3): 1551-1557.
[10] MACKEY D A, OOSTRA R J, ROSENBERG T, et al. Primary pathogenic mtDNA mutations in multigeneration pedigrees with Leber hereditary optic neuropathy[J]. Am J Hum Genet, 1996, 59(2): 481-485.
[11] LIANG M, JIANG P, LI F, et al. Frequency and spectrum of mitochondrial ND6 mutations in 1218 Han Chinese subjects with Leber's hereditary optic neuropathy[J]. Invest Ophthalmol Vis Sci, 2014, 55(3): 1321-1331.
[12] JIA X, LI S, XIAO X, et al. Molecular epidemiology of mtDNA mutations in 903 Chinese families suspected with Leber hereditary optic neuropathy[J]. J Hum Genet, 2006, 51(10): 851-856.
[13] 郭莉, 贾小云, 肖学珊, 等. 原因不明的视神经炎线粒体DNA突变检测[J]. 中华眼底病杂志, 2000, 16(2): 78-79.
[14] LE MARCHAND L, LUM-JONES A, SALTZMAN B, et al. Feasibility of collecting buccal cell DNA by mail in a cohort study[J]. Cancer Epidemiol Biomarkers Prev, 2001, 10(6): 701-703.
[15] HARTY L C, SHIELDS P G, WINN D M, et al. Self-collection of oral epithelial cell DNA under instruction from epidemiologic interviewers[J]. Am J Epidemiol, 2000, 151(2): 199-205.
[16] HARTY L C, GARCIA-CLOSAS M, ROTHMAN N, et al. Collection of buccal cell DNA using treated cards[J]. Cancer Epidemiol Biomarkers Prev, 2000, 9(5): 501-506.
[17] BROWN M D, TORRONI A, RECKORD C L, et al. Phylogenetic analysis of Leber's hereditary optic neuropathy mitochondrial DNA's indicates multiple independent occurrences of the common mutations[J]. Hum Mutat, 1995, 6(4): 311-325.
[18] ANDREWS R M, KUBACKA I, CHINNERY P F, et al. Reanalysis and revision of the Cambridge reference sequence for human mitochondrial DNA[J]. Nat Genet, 1999, 23(2): 147.
[19] WOISCHNIK M, MORAES C T. Pattern of organization of human mitochondrial pseudogenes in the nuclear genome[J]. Genome Res, 2002, 12(6): 885-893.
[20] GONG S, PENG Y, JIANG P, et al. A deafness-associated tRNAHis mutation alters the mitochondrial function, ROS production and membrane potential[J]. Nucleic Acids Res, 2014, 42(12): 8039-8048.
[21] LI X, FISCHEL-GHODSIAN N, SCHWARTZ F, et al. Biochemical characterization of the mitochondrial tRNASer(UCN) T7511C mutation associated with nonsyndromic deafness[J]. Nucleic Acids Res, 2004, 32(3): 867-877.
[22] WANG X, LU J, ZHU Y, et al. Mitochondrial tRNAThr G15927A mutation may modulate the phenotypic manifestation of ototoxic 12S rRNA A1555G mutation in four Chinese families[J]. Pharmacogenet Genomics, 2008, 18(12): 1059-1070.
[23] 张琼敏, 郑静, 宋攀攀, 等. 口腔拭子在药物性耳聋基因检测中的应用[J]. 温州医学院学报, 2011, 41(5): 409-412.
[24] 张琼敏, 李斯斯, 陈君, 等. 儿童氨基糖苷类药物性耳聋基因快速无创检测法的应用[J]. 温州医科大学学报, 2018, 48(11): 791-795.
[25] WOUTERS J, WEIJERMAN M E, VAN FURTH A M, et al. Prospective human leukocyte antigen, endomysium immunoglobulin A antibodies, and transglutaminase antibodies testing for celiac disease in children with Down syndrome[J]. J Pediatr, 2009, 154(2): 239-242.
[26] ADRIAANSE M P, VREUGDENHIL A C, VASTMANS V, et al. Human leukocyte antigen typing using buccal swabs as accurate and non-invasive substitute for venipuncture in children at risk for celiac disease[J]. J Gastroenterol Hepatol, 2016, 31(10): 1711-1716.
[27] BONAMICO M, NENNA R, MONTUORI M, et al. First salivary screening of celiac disease by detection of anti-transglutaminase autoantibody radioimmunoassay in 5000 Italian primary schoolchildren[J]. J Pediatr Gastroenterol Nutr, 2011, 52(1): 17-20.