D'Argenio Valeria, Esposito Maria Valeria, Telese Antonella, Precone Vincenza, Starnone Flavio, Nunziato Marcella, Cantiello Piergiuseppe, Iorio Mariangela, Evangelista Eloisa, D'Aiuto Massimiliano, Calabrese Alessandra, Frisso Giulia, D'Aiuto Giuseppe, Salvatore Francesco
CEINGE-Biotecnologie Avanzate, via G. Salvatore 486, 80145 Naples, Italy; Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Via Pansini 5, 80131 Naples, Italy.
CEINGE-Biotecnologie Avanzate, via G. Salvatore 486, 80145 Naples, Italy.
Clin Chim Acta. 2015 Jun 15;446:221-5. doi: 10.1016/j.cca.2015.03.045. Epub 2015 Apr 17.
Accurate and sensitive detection of BRCA1/2 germ-line mutations is crucial for the clinical management of women affected by breast cancer, for prevention and, notably, also for the identification of at-risk healthy relatives. The most widely used methods for BRCA1/2 molecular analysis are Sanger sequencing, and denaturing high performance liquid chromatography (dHPLC) followed by the Sanger method. However, recent findings suggest that next-generation sequencing (NGS)-based approaches may be an efficient tool for diagnostic purposes. In this context, we evaluated the effectiveness of NGS for BRCA gene analysis compared with dHPLC/Sanger sequencing.
Seventy women were screened for BRCA1/2 mutations by both dHPLC/Sanger sequencing and NGS, and the data were analyzed using a bioinformatic pipeline.
Sequence data analysis showed that NGS is more sensitive in detecting BRCA1/2 variants than the conventional procedure, namely, dHPLC/Sanger.
Next-generation sequencing is more sensitive, faster, easier to use and less expensive than the conventional Sanger method. Consequently, it is a reliable procedure for the routine molecular screening of the BRCA1/2 genes.
准确且灵敏地检测BRCA1/2种系突变对于乳腺癌女性患者的临床管理、预防，尤其是对于识别有患病风险的健康亲属而言至关重要。BRCA1/2分子分析最广泛使用的方法是桑格测序法，以及变性高效液相色谱法（dHPLC）随后再采用桑格法。然而，最近的研究结果表明，基于新一代测序（NGS）的方法可能是一种用于诊断目的的有效工具。在此背景下，我们评估了与dHPLC/桑格测序相比，NGS用于BRCA基因分析的有效性。
通过dHPLC/桑格测序和NGS对70名女性进行BRCA1/2突变筛查，并使用生物信息学流程对数据进行分析。
序列数据分析表明，在检测BRCA1/2变异方面，NGS比传统方法（即dHPLC/桑格法）更灵敏。
新一代测序比传统的桑格法更灵敏、更快、更易于使用且成本更低。因此，它是BRCA1/2基因常规分子筛查的可靠方法。
