Characterization of mitochondrial DNA heteroplasmy using a parallel sequencing system
Characterization of mitochondrial DNA heteroplasmy using a parallel sequencing system
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DOI：
10.2144/000113389
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发表时间：
2010-04-01
期刊：
影响因子：
2.7
通讯作者：
Huang, Taosheng
中科院分区：
文献类型：
--
作者：
Tang, Sha;Huang, Taosheng
Characterization of human mitochondrial genome sequences is important for the molecular diagnosis of mitochondrial diseases, especially in samples with a low level of mitochondrial DNA (mtDNA) heteroplasmy (>= 5%). Currently, no single methodology can simultaneously determine complete mtDNA sequences, identify mitochondrial genome wide heteroplasmies, and quantify mtDNA heteroplasmy levels. The deep sampling inherent in "next-generation" sequencing approaches should enable the efficient detection of low-level DNA heteroplasmies and address this need. Herein, we used the Illumina Genome Analyzer to re-sequence human mtDNA samples from two subjects that were combined at five different ratios (1:99, 5:95, 10:90,20:80, and 50:50). We assessed the sensitivity, specificity, and accuracy of this system, and our results show that mtDNA heteroplasmies >= 5% were detected 100% of the time with virtually no false positives and that the estimates of mtDNA heteroplasmy levels were remarkably close to the theoretical values (correlation coefficient = 0.96). Therefore, parallel sequencing provides a simple, high-throughput, and cost-effective platform for mitochondrial genome sequencing with sensitivity and specificity for mcDNA hetcroplasmy detection.
