Improved detection of the G1528C mutation in LCHAD deficiency
Improved detection of the G1528C mutation in LCHAD deficiency
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DOI：
10.1006/bmme.1996.0031
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发表时间：
1996-06-01
期刊：
影响因子：
--
通讯作者：
Roe, CR
中科院分区：
文献类型：
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作者：
Ding, JH;Yang, BZ;Roe, CR
长链3-羟基酰辅酶A脱氢酶(LCHAD)缺乏症是一种脂肪酸氧化的常染色体隐性遗传性疾病，临床上以骨骼肌病、Reye样综合征或原因不明的婴儿猝死为特征。最近报道了一种常见的突变G1528C。为了避免“假基因”带来的非特异性扩增和潜在的并发症，我们开发了一种套式聚合酶链式反应/PstI消化方法。在这里，我们报告了另外11例LCHAD缺乏症患者的突变研究。用套式聚合酶链式反应扩增基因组DNA片段(117bp)，经PstI酶切后，在12%聚丙烯酰胺凝胶上进行电泳。4例患者G1528C突变为纯合子，7例为复合杂合子，表明存在显著的遗传异质性。在所有孤立性LCHAD缺乏症患者中，至少在一个等位基因上发现了G1528C突变，这表明G1528C突变是该病的一个很好的标记。这种DNA检测与串联质谱学体外探针分析相结合，很容易识别G1528C复合杂合子家系中的患病个体和携带者。(C)1996年学术出版社。
Long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency, an autosomal recessive disorder of fatty-acid oxidation, is clinically characterized by skeletal myopathy, Reye-like syndrome, or sudden unexplained infant death. A common mutation, G1528C, has recently been reported. To avoid nonspecific amplification from a ''pseudogene'' and potential complications, we have developed a nested PCR/PstI digestion method. Here, we report mutation studies in 11 additional unrelated patients with LCHAD deficiency. Genomic DNA fragments (117 bp) were amplified by the nested PCR, digested with PstI, and subjected to electrophoresis on 12% polyacrylamide gel. Four patients were found to be homozygous for the G1528C mutation; 7 patients were compound heterozygous, indicating significant genetic heterogeneity. The G1528C mutation has been found on at least one allele in all patients with isolated LCHAD deficiency, suggesting that it is an excellent marker for this disease. This DNA test combined with tandem mass-spectrometric in vitro probe analysis easily identifies affected individuals and carriers in families which are compound heterozygous for G1528C. (C) 1996 Academic Press, Inc.