Direct detection of a BRAF mutation in total RNA from melanoma cells using cantilever arrays
Direct detection of a BRAF mutation in total RNA from melanoma cells using cantilever arrays
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DOI：
10.1038/nnano.2012.263
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发表时间：
2013-02-01
影响因子：
38.3
通讯作者：
Gerber, Ch
中科院分区：
文献类型：
--
作者：
Huber, F.;Lang, H. P.;Gerber, Ch
恶性黑色素瘤是最致命的皮肤癌形式，其特征在于BRAF基因的主要突变（1-3）。靶向携带这种突变的肿瘤的药物最近已进入临床（4-7）。因此，常规筛查患者的这种基因突变，以确定他们是否可以从这种类型的治疗中获益。目前突变筛查的金标准使用实时聚合酶链反应和测序方法（8）。在这里，我们表明，基于微悬臂梁阵列的测定可以检测突变nanomechanically从黑色素瘤细胞分离的总RNA样品中没有扩增。该测定基于BRAF特异性寡核苷酸探针。我们检测到突变BRAF在浓度为500 pM的野生型序列的50倍过量。该方法能够区分携带突变的黑色素瘤细胞和野生型细胞，只需20 ng μ l（-1）的RNA材料，而无需事先进行PCR扩增和使用标记。
Malignant melanoma, the deadliest form of skin cancer, is characterized by a predominant mutation in the BRAF gene(1-3). Drugs that target tumours carrying this mutation have recently entered the clinic(4-7). Accordingly, patients are routinely screened for mutations in this gene to determine whether they can benefit from this type of treatment. The current gold standard for mutation screening uses real-time polymerase chain reaction and sequencing methods(8). Here we show that an assay based on microcantilever arrays can detect the mutation nanomechanically without amplification in total RNA samples isolated from melanoma cells. The assay is based on a BRAF-specific oligonucleotide probe. We detected mutant BRAF at a concentration of 500 pM in a 50-fold excess of the wild-type sequence. The method was able to distinguish melanoma cells carrying the mutation from wild-type cells using as little as 20 ng mu l(-1) of RNA material, without prior PCR amplification and use of labels.