Yang Ju-hua, Tong Yi, Li Bao-hua, Chen Yi-kai
The First Affiliated Hospital, Fujian Medical University, Fuzhou 350004, China.
Zhonghua Yan Ke Za Zhi. 2005 Mar;41(3):243-5.
Rapid Genetic Screening of Leber's hereditary optic neuropathy (LHON) with mtDNA G11778A mutation by allele-specific polymerase chain reaction (AS-PCR) with whole blood.
Whole blood with anticoagulant was used as a template of AS-PCR for the analysis of LHON with mtDNA G11778A point mutation. The amplified DNA fragment was directly observed by electrophoretogram with ethidium bromide stained.
The accuracy was 100% by using this method in 24 blood samples tested, and the specific of PCR of which used whole blood as template was better than one of the purified mtDNA. The reliability of the method for screening of LHON with mtDNA G11778A mutation was checked by double-blind test in 22 blood samples.
This method does not need purified DNA from blood and only required one step of PCR. Thus, it is very simple, rapid and accurate for the clinical genetic screening of LHON with mtDNA G11778A point mutation.
采用等位基因特异性聚合酶链反应（AS-PCR）技术，利用全血对伴有线粒体DNA G11778A突变的Leber遗传性视神经病变（LHON）进行快速基因筛查。
以抗凝全血作为AS-PCR的模板，用于分析伴有线粒体DNA G11778A点突变的LHON。扩增的DNA片段经溴化乙锭染色后通过电泳图谱直接观察。
用该方法检测24份血样，准确率达100%，以全血为模板的PCR特异性优于以纯化线粒体DNA为模板的PCR。通过对22份血样进行双盲试验，检验了该方法筛查伴有线粒体DNA G11778A突变的LHON的可靠性。
该方法无需从血液中纯化DNA，仅需一步PCR。因此，对于伴有线粒体DNA G11778A点突变的LHON的临床基因筛查，该方法非常简单、快速且准确。
