Coppin Lucie, Grutzmacher Claudine, Crépin Michel, Destailleur Evelyne, Giraud Sophie, Cardot-Bauters Catherine, Porchet Nicole, Pigny Pascal
1] Laboratoire de biochimie 'hormonologie, métabolisme-nutrition, oncologie', Centre de Biologie Pathologie, CHRU de Lille, Lille Cedex, France [2] INSERM, U837, Team 5, Lille Cedex, France.
Laboratoire de biochimie 'hormonologie, métabolisme-nutrition, oncologie', Centre de Biologie Pathologie, CHRU de Lille, Lille Cedex, France.
Eur J Hum Genet. 2014 Sep;22(9):1149-52. doi: 10.1038/ejhg.2013.279. Epub 2013 Dec 4.
The identification of Von Hippel-Lindau (VHL) mosaic mutations by conventional Sanger sequencing requires a labour-intensive enrichment step, thus explaining that mosaicism occurrence is underestimated in patients. Nowadays, it is possible to detect mutation in cell sub-populations by next-generation sequencing (NGS). Here, we described a diagnosis strategy using NGS with high coverage in a series of eight patients who were negative for a VHL abnormality by Sanger sequencing and deletion search. In two patients, a mosaic mutation in VHL was detected by NGS. One patient with a 5.7% mutated allele frequency had a severe phenotype and an early disease onset. In conclusion, clinical NGS in an hospital molecular oncogenetics laboratory is an efficient tool to identify VHL mosaic mutation. Its use may improve patient monitoring and genetic counseling.
通过传统桑格测序法鉴定冯·希佩尔-林道(VHL)镶嵌突变需要进行劳动强度大的富集步骤,这就解释了为何患者中镶嵌现象的发生率被低估。如今,通过新一代测序(NGS)能够检测细胞亚群中的突变。在此,我们描述了一种诊断策略,该策略在一系列8例经桑格测序和缺失搜索未发现VHL异常的患者中使用了高覆盖度的NGS。在两名患者中,通过NGS检测到了VHL的镶嵌突变。一名等位基因突变频率为5.7%的患者具有严重的表型且疾病发病较早。总之,医院分子肿瘤遗传学实验室中的临床NGS是鉴定VHL镶嵌突变的有效工具。其应用可能会改善患者监测和遗传咨询。