Study on real-time probe technique detecting mtDNA 11778 point mutation in Leber hereditary optic neuropathy patients
10.3760/cma.j.issn.2095-0160.2013.02.014
- VernacularTitle:荧光探针法检测Leber遗传性视神经病变11778位点突变的研究
- Author:
Yan-min, ZHANG
;
Dan, SONG
;
Hui-juan, AN
;
Kang, CHEN
;
Zhi-li, WANG
;
Yu-zhou, BAO
;
Zhao-xia, ZHAO
- Publication Type:Journal Article
- Keywords:
Leber hereditary optic neuropathy;
Mitochondrial DNA;
Real-time fluorescence quantitative polymerase chain reaction;
Point mutation
- From:
Chinese Journal of Experimental Ophthalmology
2013;(2):164-167
- CountryChina
- Language:Chinese
-
Abstract:
Background Although Leber hereditary optic neuropathy (LHON) and optic neuritis have different causes and managements,their clinical manifestations are difficult to be distinguished.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR) is a high flux,simple,rapid and specific detecting technology,so establishing a specific diagnosis method of LHON with RTFQ-PCR has a practical significance.Objective Purpose of the present study was to establish a real-time Taqman probe for mitochondrial DNA (mtDNA)11778G＞A mutation in LHON patients.Methods Primers and Taqman probe for mtDNA 11778G＞A mutation were designed based on mtDNA complete geneme.Eighty-four patients with LHON were selected from the LHON DNA bank of Molecular Biology Laboratory,Henan Eye Institute,and 40 normal physical examinees aged 18-20 years were from Henan People's Hospital.2 ml of periphery blood was collected from each individual.Based on the double-blindness principle,mtDNA 11778G＞A mutation was tested by both Taqman probe and sequencing to check the reliability of real-time Taqman probe.Results The mtDNA 11778G＞A mutation was found in 23 out of 84 patients,and 61 showed a negative result by the technique of real-time Taqman probe.The Ct values of 23 patients with mtDNA 11778G＞A mutation were 22.993 ±0.708,but those of 5 normal controls were 0.These findings showed a consistent rate of 100％ with the sequencing results.In addition,both the false positive rate and the false negative rate were zero.Conclusions Real-time Taqman probe technique is an accurate,convenient,sensitive,specific and intuitionistic method for the diagnosis of mtDNA 11778G＞A mutation in LHON patients.It is feasible and suitable to screen the LHON patients with mtDNA 11778G＞A mutation in a large scale.
