Vrabelova Slavka, Letocha Ondrej, Borsky Marek, Kozak Libor
Center of Molecular Biology and Gene Therapy, University Hospital Brno, Cernopolni 9, 625 00 Brno, Czech Republic.
Mol Genet Metab. 2005 Sep-Oct;86(1-2):277-85. doi: 10.1016/j.ymgme.2005.05.004. Epub 2005 Jun 20.
Wilson disease (WD) is an autosomal recessive disorder of copper transport. WD patients are presenting with a wide range of heterogeneous clinical syndromes including hepatic, neurological, or psychiatric presentations. The disease is caused by mutations in the ATP7B gene. This study presents the results of comprehensive mutation analysis in 227 WD patients from 200 unrelated families (173 from Czech Republic and 27 from Slovakia). More than 80% of all mutant alleles were identified, using a combination of PCR/RFLP, DGGE, TTGE, DHPLC, and sequencing. A total of 40 different mutations and 18 polymorphisms were detected on 400 independent mutant chromosomes. The most common molecular defect was H1069Q (57% of all 400 studied alleles). Each of the other 39 mutations was present in no more than 4% of WD alleles and 23 mutations were found in only one WD allele each (0.25%). Thirteen novel mutations were identified, including seven missense mutations (L641S, T737R, D918E, T1033S, G1111D, D1271N, and G1355C), four small deletions (19_20delCA, 1518_1522del5, 3140delA, and 3794_3803del10), and two splice-site mutations (2446-2A>G, 2865+1G>A). We did not find a significant correlation between H1069Q homozygosity and age of onset, and clinical and biochemical manifestation. Our data provide evidence that the H1069Q mutation-the most common molecular defect of the ATP7B gene in the Caucasian population-originates from Central/Eastern Europe. Screening of five prevalent mutations is predicted to reveal 70% of all mutant alleles presented in WD patients. This will provide a good starting point for early clinical classification of WD in our population.
威尔逊病（WD）是一种常染色体隐性铜转运障碍疾病。WD患者表现出广泛的异质性临床综合征，包括肝脏、神经或精神方面的症状。该疾病由ATP7B基因突变引起。本研究展示了对来自200个无亲缘关系家庭（173个来自捷克共和国，27个来自斯洛伐克）的227例WD患者进行全面突变分析的结果。通过聚合酶链反应/限制性片段长度多态性分析（PCR/RFLP）、变性梯度凝胶电泳（DGGE）、温度梯度凝胶电泳（TTGE）、变性高效液相色谱（DHPLC）和测序等方法的组合，鉴定出了超过80%的所有突变等位基因。在400条独立的突变染色体上共检测到40种不同的突变和18种多态性。最常见的分子缺陷是H1069Q（在所有400个研究等位基因中占57%）。其他39种突变中的每一种在WD等位基因中的出现频率均不超过4%，且有23种突变仅在一个WD等位基因中被发现（0.25%）。鉴定出了13种新突变，包括7种错义突变（L641S、T737R、D918E、T1033S、G1111D、D1271N和G1355C）、4种小缺失（19_20delCA、1518_1522del5、3140delA和3794_3803del）以及2种剪接位点突变（2446-2A>G、2865+1G>A）。我们未发现H1069Q纯合性与发病年龄、临床及生化表现之间存在显著相关性。我们的数据表明，H1069Q突变——白种人群中ATP7B基因最常见的分子缺陷——起源于中欧/东欧。预计对5种常见突变进行筛查可揭示WD患者中所有突变等位基因的70%。这将为我国人群中WD的早期临床分类提供一个良好的起点。
