Screening for mutations in BRCA1 and BRCA2 genes by measuring the acoustic ratio with QCM
Screening for mutations in BRCA1 and BRCA2 genes by measuring the acoustic ratio with QCM
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DOI：
10.1039/c3ay41143e
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发表时间：
2014
影响因子：
3.1
通讯作者：
G. Papadakis;E. Gizeli
中科院分区：
文献类型：
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作者：
G. Papadakis;E. Gizeli
肿瘤抑制基因BRCA 1和BRCA 2突变的筛查对于乳腺癌和卵巢癌的预防具有重要意义。我们描述了一种基于声波装置的突变筛选和检测方法。特别是，我们检测四个突变位于BRCA 1和BRCA 2基因使用石英晶体微天平技术。该检测基于PCR扩增和限制性酶切消化后产生的与装置表面结合的双链DNA分子的耗散与频率变化的声学比（ΔD/ΔF）的测量;声学比已被证明是附着分子的固有粘度的量度，而固有粘度又取决于dsDNA的大小。该方法的新特征是缺少杂交步骤，DNA分子的长度而不是质量的无标记感测，以及直接检测消化的DNA产物而无需事先纯化。该方法是通用的，简单的，能够检测单碱基突变长基因组重排;它也是合适的，适用于芯片实验室的概念。
Screening for mutations in the tumor-suppressor genes BRCA1 and BRCA2 is of great importance for breast and ovarian cancer prevention. We describe a methodology for mutation screening and detection based on acoustic wave devices. In particular, we detect four mutations located in BRCA1 and BRCA2 genes using the quartz crystal microbalance technique. The detection is based on measurements of the acoustic ratio of dissipation versus frequency change (ΔD/ΔF) of double-stranded DNA molecules bound to the device surface that are produced after PCR amplification and restriction digestion; the acoustic ratio has been shown to be a measure of the intrinsic viscosity of the attached molecules, which, in turn, depends on the size of the dsDNAs. Novel features of this approach are the lack of a hybridization step, the label free sensing of the length, rather than mass, of the DNA molecules and the direct detection of the digested DNA products without prior purification. The method is generic, simple and capable of detecting single base mutations to long genomic rearrangements; it is also suitable and applicable to a Lab-on-a-chip concept.