Detection and quantitation of mitochondrial G11778A mutation of LHON syndrome in a Vietnamese patient with tentatively diagnosed mitochondrial disease
Main Article Content
Abstract
G11778A mutation in the ND4 gene of human mitochondrial genome accounts for 50-70% of LHON syndrome. In this study, we set up real-time PCR using fluorescent Taqman probe with locked nucleic acid nucleotide (LNA) for detection and quantitation of mitochondrial genome mutation G11778A. The real-time PCR showed a linear correlation between logarithm of target gene copy number and threshold value (Ct) with a high regression value R2 = 0.999. By using PCR-RFLP in combination of real-time PCR, we found a 7.5 month girl patient carrying G11778A mutation. The mutation was present at 2.71±0.12% of heteroplasmy. whereas her parents did not carry this mutation. This is the first case with the mutation G11778A found in Vietnam.
Keywords:
G11778A mutation, LHON syndrome, PCR-RFLP, Real-time PCR, LNA probe
References
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[3] Mashima Y., Leber’s hereditary optic neuropathy, Nippon Rinsho 60 4 (2002) 282.
[4] Robinson B., H., Lactic acidemia and mitochondrial disease, Mol Genet Metab 89 (2006) 3.
[5] Jia X., Li S., Xiao X., Guo X., Zhang Q., Molecular epidemiology of mtDNA mutations in 903 Chinese families suspected with Leber hereditary optic neuropathy, J Hum Genet 51 (2006) 851.
[6] Mackey D. A., Oostra R. J., Rosenberg T., Nikoskelainen E., Bronte-Stewart J., Poulton J., Harding A. E., Govan G., Bolhuis P. A., Norby S., Primary pathogenic mtDNA mutations in multigeneration pedigrees with Leber hereditary optic neuropathy, Am J Hum Genet 59 (1996) 481.
[7] Mashima Y., Nagano M., Funayama T., Zhang Q., Egashira T., Kudho J., Shimizu N., Oguchi Y., Rapid quantification of the heteroplasmy of mutant mitochondrial DNAs in Leber’s hereditary optic neuropathy using the Invader technology, Clin Biochem 37 (2003) 268.
[8] Mashima Y., Yamada K., Wakakura M., Kigasawa K., Kudoh J., Shimizu N., Oguchi Y., Spectrum of pathogenic mitochondrial DNA mutations and clinical features in Japanese families with Leber’s hereditary optic neuropathy, Curr Eye Res 17 (1998) 403.
[9] Chinnery P. F., Andrews R.M., Turnbull D.M., Howell N.N., Leber hereditary optic neuropathy: Does heteroplasmy influence the inheritance and expression of the G11778A mitochondrial DNA mutation? Am J Med Genet 98 (2001) 235.
[10] Wang J. Y., Gu Y. S., Wang J., Tong Y., Wang Y., Shao J. B., Qi M., MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR, J Zhej Univ Sci B 9 8 (2008) 610.
[11] Nguyễn Văn Minh, Phùng Bảo Khánh, Trương Thị Huệ, Cao Vũ Hùng, Phạm Vân Anh, Nguyễn Thị Hồng Loan, Phan Tuấn Nghĩa (2015) Sàng lọc đột biến G3460A, G11778A và phát hiện đột biến mất đoạn 6 bp mới trên hệ gen ty thể bệnh nhân thần kinh cơ Việt nam, Tạp chí Công nghệ sinh học 13 (2) 213.
[12] Truong T. H., Nguyen T. V. A., Nguyen T. H. L., Pham V. A., Phan T. N., Sensitive quantification of mitochondrial mutation using new Taqman probes, Cent Eur J Med 9 6 (2014) 839.
[13] Smith K., H., Johns D., R., Heher K. L., Miller N. R., Heteroplasmy in Leber’s hereditary optic neuropathy, Arch Ophthalmol 111 (1993) 1486.