Abernathy C R, Rasmussen S A, Stalker H J, Zori R, Driscoll D J, Williams C A, Kousseff B G, Wallace M R
Department of Pediatrics, University of Florida, Gainesville 32610-0296, USA.
Hum Mutat. 1997;9(6):548-54. doi: 10.1002/(SICI)1098-1004(1997)9:6<548::AID-HUMU8>3.0.CO;2-Y.
Neurofibromatosis type 1 (NF1) is a common autosomal dominant disorder characterized predominantly by neurofibromas, café-au-lait spots, and Lisch nodules. The disease is caused by disruptive mutations of the large NF1 gene, with half of cases caused by new mutation. Less than 100 constitutional mutations have thus far been published, ranging from very large deletions to point mutations. We have pursued NF1 mutation analysis by heteroduplex analysis (HDA) and single-strand conformational polymorphism analysis (SSCP) of individual exons. We streamlined these techniques to eliminate the use of radioactivity, to apply both methods to the same PCR product, and to multiplex samples in gels. Applied simultaneously to a set of 67 unrelated NF1 patients, HDA and SSCP have thus far identified 26 mutations and/or variants in 45 of the 59 exons tested. Disease-causing mutations were found in 19% (13/67) of cases studied. Both techniques detected a variety of mutations including splice mutations, insertions, deletions, and point changes, with some overlap in the ability of each method to detect variants.
1型神经纤维瘤病（NF1）是一种常见的常染色体显性疾病，主要特征为神经纤维瘤、咖啡斑和Lisch结节。该疾病由大型NF1基因突变引起，其中一半病例由新发突变导致。迄今为止，已发表的不到100个胚系突变，范围从非常大的缺失到点突变。我们通过对单个外显子进行异源双链分析（HDA）和单链构象多态性分析（SSCP）来进行NF1突变分析。我们简化了这些技术，以消除放射性的使用，将两种方法应用于同一PCR产物，并在凝胶中对样本进行多重分析。将HDA和SSCP同时应用于一组67名无亲缘关系的NF1患者，迄今为止，在检测的59个外显子中的45个中鉴定出26个突变和/或变异。在19%（13/67）的研究病例中发现了致病突变。两种技术都检测到了多种突变，包括剪接突变、插入、缺失和点变化，每种方法检测变异的能力有一些重叠。
