Application and evaluation of denaturing HPLC for molecular genetic analysis in tuberous sclerosis
Application and evaluation of denaturing HPLC for molecular genetic analysis in tuberous sclerosis
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变性高效液相色谱在结节性硬化症分子遗传学分析中的应用及评价
DOI：
10.1007/s004390000316
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发表时间：
2000
期刊：
影响因子：
5.3
通讯作者：
J. Cheadle
中科院分区：
文献类型：
--
作者：
Alistair Jones;J. Sampson;B. Hoogendoorn;D. Cohen;J. Cheadle
抽象的。结节性硬化症（TSC）是一种常染色体显性遗传疾病，其特征是多个组织和器官发生错构瘤。TSC表现出基因座异质性，9 q34（TSC 1）和16p13.3（TSC 2）的基因分别具有21和41个编码外显子。这两个基因座的突变谱很广，以前的研究表明，60%-70%的病例是散发性的，代表新的突变。我们已经格式化变性高效液相色谱（DHPLC）快速筛选所有编码外显子的TSC 1和TSC 2。DHPLC分析在150例无关病例中检测到103例（68%）可能的致病突变，而单链构象多态性分析（SSCP）和传统异源双链分析（HA）分别为92/150（61%）和87/150（58%）。确定每个外显子筛查程序的资本、耗材和劳动力成本。每个患者样本的估计成本取决于吞吐量，特别是对于DHPLC，其中高比例的成本是固定的，并且对于DHPLC，SSCP和HA分别为257英镑，216英镑和242英镑，假设每年的吞吐量为252个样本，或354英镑，233英镑和259英镑，假设每年的吞吐量为126个样本。DHPLC的优势，增加了灵敏度和降低劳动力成本相比，更传统的方法，外显子筛选，但除非昂贵的DHPLC设备被有效地利用了很高比例的时间，总成本略高。
Abstract. Tuberous sclerosis (TSC) is an autosomal dominant disorder characterised by the development of hamartomas in multiple tissues and organs. TSC exhibits locus heterogeneity with genes at 9q34 (TSC1) and 16p13.3 (TSC2) that have 21 and 41 coding exons, respectively. The mutational spectrum at both loci is wide and previous studies have shown that 60%–70% of cases are sporadic and represent new mutations. We have formatted denaturing high performance liquid chromatography (DHPLC) for rapid screening of all coding exons of TSC1 and TSC2. DHPLC analysis detected likely disease-causing mutations in 103 of 150 unrelated cases (68%), compared with 92/150 (61%) and 87/150 (58%) for single-strand conformation polymorphism analysis (SSCP) and conventional heteroduplex analysis (HA), respectively. Capital, consumable and labour costs were determined for each exon screening procedure. Estimated costs per patient sample depended on throughput, particularly for DHPLC, where a high proportion of costs are fixed, and were £257, £216 and £242 for DHPLC, SSCP and HA, respectively, assuming a throughput of 252 samples per year, or £354, £233 and £259, assuming a throughput of 126 samples per year. DHPLC had the advantages of increased sensitivity and reduced labour costs when compared with more traditional approaches to exon screening but, unless expensive DHPLC equipment is being efficiently utilised for a very high proportion of the time available, overall costs are slightly higher.
等位基因丢失在结节性硬化症肾脏病变中很常见，但在脑病变中很少见。
影响因子：
9.8
作者：
Henske,EP;Scheithauer,BW;Short,MP;Wollmann,R;Nahmias,J;Hornigold,N;vanSlegtenhorst,M;Welsh,CT;Kwiatkowski,DJ
通讯作者：
Kwiatkowski,DJ