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Journal of Zhejiang University SCIENCE B 2008 Vol.9 No.8 P.610~615


MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR

Author(s):  Jian-yong WANG, Yang-shun GU, Jing WANG, Yi TONG, Ying WANG, Jun-bing SHAO, Ming QI

Affiliation(s):  Department of Ophthalmology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China; more

Corresponding email(s):   Guyangshun_1@hotmail.com

Key Words:  Leber&rsquo, s hereditary optic neuropathy (LHON), Mitochondrial DNA (mtDNA), MtDNA11778 mutation, Minor groove binder (MGB) probe, Real-time polymerase chain reaction (PCR)

Jian-yong WANG, Yang-shun GU, Jing WANG, Yi TONG, Ying WANG, Jun-bing SHAO, Ming QI. MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR[J]. Journal of Zhejiang University Science B, 2008, 9(8): 610~615.

@article{title="MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR",
author="Jian-yong WANG, Yang-shun GU, Jing WANG, Yi TONG, Ying WANG, Jun-bing SHAO, Ming QI",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR
%A Jian-yong WANG
%A Yang-shun GU
%A Jing WANG
%A Ying WANG
%A Jun-bing SHAO
%A Ming QI
%J Journal of Zhejiang University SCIENCE B
%V 9
%N 8
%P 610~615
%@ 1673-1581
%D 2008
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0820058

T1 - MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR
A1 - Jian-yong WANG
A1 - Yang-shun GU
A1 - Jing WANG
A1 - Yi TONG
A1 - Ying WANG
A1 - Jun-bing SHAO
A1 - Ming QI
J0 - Journal of Zhejiang University Science B
VL - 9
IS - 8
SP - 610
EP - 615
%@ 1673-1581
Y1 - 2008
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0820058

Objective: leber&rsquo;s hereditary optic neuropathy (LHON) is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA (mtDNA). Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing. This study aims to develop a minor groove binder (MGB) probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction (PCR). Methods: Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation, with 20 normal individuals as a control group at the same time. A real-time PCR involving two MGB probes was used to detect the mtDNA11778 mutation and heteroplasmy. A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones. Results: All 48 LHON patients and their maternal relatives were positive for mtDNA11778 mutation in our assay, 27 heteroplasmic and 21 homoplasmic. Eighteen cases did not show an occurrence of the disease, while 9 developed the disease among the 27 heteroplasmic mutation cases. Eleven did not show an occurrence of the disease, while 10 cases developed the disease among 21 homoplasmic mutation cases. There was a significant difference in the incidence between the heteroplasmic and the homoplasmic mutation types. The time needed for running a real-time PCR assay was only 80 min. Conclusion: This real-time PCR assay is a rapid, reliable method for mtDNA mutation detection as well as heteroplasmy quantification. Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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