Division of Molecular Diagnostics, Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Clin Chim Acta. 2010 May 2;411(9-10):690-9. doi: 10.1016/j.cca.2010.01.033. Epub 2010 Feb 6.
Multiple acyl-CoA dehydrogenase deficiency (MADD) or gluaric aciduria type II is an autosomal recessive disease caused by defects in mitochondrial electron transfer system and metabolism of fatty acid. Recently, ETFDH mutations were reported to be major causes of riboflavin-responsive MADD. The present study is aimed at screening ETFDH mutations.
High resolution melting (HRM) analysis was performed to screen ETFDH mutations. Genomic DNA was extracted from peripheral blood samples of the 9 patients with MADD and normal controls. Total 13 exons of ETFDH were screened by HRM analysis. The results were subsequently confirmed by direct DNA sequencing.
This diagnostic strategy proved to be feasible in detecting 3 known (c.250G>A, c380T>A, c.524G>T) and 1 novel (c.1831G>A) ETFDH mutations. Each mutation could be readily and accurately identified in the difference plot curves. We estimated the carrier frequency of the hotspot mutation, c.250G>A, in the Taiwanese population to be 1:125 (0.8%).
HRM analysis can be successfully applied to screen ETFDH mutations. Since riboflavin-responsive MADD is often treatable, especially with mutations in ETFDH, identifying ETFDH mutations is crucial for these patients.
多种酰基辅酶 A 脱氢酶缺乏症（MADD）或戊二酸尿症 II 型是一种常染色体隐性疾病，由线粒体电子传递系统和脂肪酸代谢缺陷引起。最近，ETFDH 突变被报道为反应性核黄素 MADD 的主要原因。本研究旨在筛选 ETFDH 突变。
高分辨率熔解（HRM）分析用于筛选 ETFDH 突变。从 9 例 MADD 患者和正常对照的外周血样本中提取基因组 DNA。通过 HRM 分析筛选 ETFDH 的 13 个外显子。结果随后通过直接 DNA 测序确认。
该诊断策略在检测 3 种已知（c.250G>A、c380T>A、c.524G>T）和 1 种新（c.1831G>A）的 ETFDH 突变时被证明是可行的。每个突变都可以在差异图谱曲线中轻易且准确地识别。我们估计热点突变 c.250G>A 在台湾人群中的携带频率为 1:125（0.8%）。
HRM 分析可成功用于筛选 ETFDH 突变。由于反应性核黄素 MADD 通常是可治疗的，特别是与 ETFDH 突变有关，因此确定 ETFDH 突变对于这些患者至关重要。
