A Simple Oligonucleotide Biochip Capable of Rapidly Detecting Known Mitochondrial DNA Mutations in Chinese Patients with Leber’S Hereditary Optic Neuropathy (LHON)

• DOI: 10.1155/2011/340723
• Publication Date: 2013-06-13T07:43:35Z
• Authors: Wei-Dong Du; Gang Chen; Hui-Min Cao; Qing-Hui Jin; Rong-Feng Liao; Xiang-Cheng He; Da-Ben Chen; Shu-Ren Huang; Hui Zhao; Yong-Mei Lv; Hua-Yang Tang; Xian-Fa Tang; Yong-Qing Wang; Song Sun; Jian-Long Zhao; Xue-Jun Zhang
• Keywords: Biochip; Heteroplasmy; Multiplex; genomic DNA

Leber's hereditary optic neuropathy (LHON) is a maternally transmitted disease. Clinically, no efficient assay protocols have been available. In this study, we aimed to develop an oligonucleotide biochip specialized for detection of known base substitution mutations in mitochondrial DNA causing LHON and investigate frequencies relevant variants Anhui region China. Thirty-two pairs probes matched with the potentially linked were covalently immobilized. Cy5-lablled targets amplified from blood samples by multiplex PCR method. Two kinds primary 11778 G > A 14484 T C six confirmed patients interrogated validate format. Further, fourteen Chinese pedigrees twenty-five unrelated healthy individuals investigated biochip, direct sequencing pyrosequencing, respectively. The was found be able efficiently discriminate homoplasmic heteroplasmic mtDNA LHON. Biochip analysis revealed that twelve eighteen symptomatic cases 14 pedigree harbored either 11778G A, 14484T or 3460G respectively, accounting 66.7%. mutation these prevalent (55.5%, 10 18 cases). 3394T co-exist one case. 13708G appeared pedigree. Smaller amount sampling reaction volume, easier target preparation, fast high-throughput main advantages over pyrosequencing. Our findings suggested are gene leading would easily implemented clinical diagnosis.