INAC, SPrAM (UMR 5819, CEA, CNRS, UJF), CEA Grenoble, 17 rue des Martyrs, F-38054 Grenoble cedex 9, France.
Biosens Bioelectron. 2012 Jan 15;31(1):554-7. doi: 10.1016/j.bios.2011.10.012. Epub 2011 Oct 17.
The possibility to detect low abundant DNA point mutations is essential for early cancer diagnosis and/or prognosis. Furthermore, in order to be less invasive, the somatic mutations are not only sought in tumor extract samples but also from body fluids or stools rendering their content even more diluted compared to the wild type sequences. In this short communication, we propose two protocols based on temperature scans or cycles for the enrichment of the mutation strands hybridized on microarrays. We predict numerically and confirm experimentally a 10-fold increase in the fraction of mutated DNA hybridized on the microarray compared to the sample content. Coupled to more standard solution phase enrichment techniques, it would be possible to lower by one order of magnitude the current detection limit with the advantage of multiple mutation detections offered by the microarray technology.
检测低丰度 DNA 点突变的可能性对于早期癌症诊断和/或预后至关重要。此外,为了减少侵入性,体细胞突变不仅在肿瘤提取样本中寻找,而且在体液或粪便中寻找,使它们的含量与野生型序列相比更加稀释。在本简讯中,我们提出了两种基于温度扫描或循环的方案,用于富集杂交在微阵列上的突变链。我们通过数值预测并通过实验证实,与样品含量相比,杂交在微阵列上的突变 DNA 分数增加了 10 倍。与更标准的溶液相富集技术相结合,有可能将当前的检测极限降低一个数量级,同时利用微阵列技术提供的多种突变检测优势。