Uusitalo Elina, Hammais Anna, Palonen Elina, Brandt Annika, Mäkelä Ville-Veikko, Kallionpää Roope, Jouhilahti Eeva-Mari, Pöyhönen Minna, Soini Juhani, Peltonen Juha, Peltonen Sirkku
Department of Cell Biology and Anatomy, University of Turku, Turku, Finland.
Acta Derm Venereol. 2014 Nov;94(6):663-6. doi: 10.2340/00015555-1843.
Neurofibromatosis type 1 syndrome (NF1) is caused by mutations in the NF1 gene. Availability of new sequencing technology prompted us to search for an alternative method for NF1 mutation analysis. Genomic DNA was isolated from saliva avoiding invasive sampling. The NF1 exons with an additional 50bp of flanking intronic sequences were captured and enriched using the SeqCap EZ Choice Library protocol. The captured DNA was sequenced with the Roche/454 GS Junior system. The mean coverages of the targeted regions were 41x and 74x in 2 separate sets of samples. An NF1 mutation was discovered in 10 out of 16 separate patient samples. Our study provides proof of principle that the sequence capture methodology combined with high-throughput sequencing is applicable to NF1 mutation analysis. Deep intronic mutations may however remain undetectable, and change at the DNA level may not predict the outcome at the mRNA or protein levels.
1型神经纤维瘤病综合征（NF1）由NF1基因突变引起。新测序技术的出现促使我们寻找一种用于NF1突变分析的替代方法。从唾液中分离基因组DNA，避免侵入性采样。使用SeqCap EZ Choice文库方案捕获并富集带有额外50bp侧翼内含子序列的NF1外显子。捕获的DNA用罗氏/454 GS Junior系统进行测序。在两组独立样本中，目标区域的平均覆盖度分别为41倍和74倍。在16份独立患者样本中的10份中发现了NF1突变。我们的研究提供了原理证明，即序列捕获方法与高通量测序相结合适用于NF1突变分析。然而，内含子深处的突变可能仍然无法检测到，而且DNA水平的变化可能无法预测mRNA或蛋白质水平的结果。
