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首页> 外文期刊>The Journal of molecular diagnostics: JMD >Rapid detection of the epidermal growth factor receptor mutation in non-small-cell lung cancer for analysis of acquired resistance using molecular beacons.
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Rapid detection of the epidermal growth factor receptor mutation in non-small-cell lung cancer for analysis of acquired resistance using molecular beacons.

机译：非小细胞肺癌中表皮生长因子受体突变的快速检测，用于使用分子信标分析获得性耐药。

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A secondary mutation (T790M) in epidermal growth factor receptor (EGFR) is a hallmark of acquired resistance to EGFR inhibitors used to treat non-small-cell lung cancer (NSCLC). Therefore, identifying the T790M mutation is crucial to guide treatment decisions. Given that DNA sequencing methods are time-consuming and insensitive, we developed and investigated the feasibility of using molecular beacons for the detection of the T790M mutation in EGFR. A molecular beacon complementary to the region of the secondary EGFR mutation (T790M) was designed and used in NSCLC samples bearing drug-sensitive and -resistant EGFR mutations. For a rapid and simple assay, we attempted to use the molecular beacon with real-time PCR and in situ fluorescence imaging. The ability of the designed molecular beacon to specifically detect the T790M mutation of EGFR was tested for samples from two patients with drug resistance and compared with conventional DNA sequencing methods. The molecular beacon successfully detected the T790M mutation in patient samples with drug resistance. The sensitivity of the molecular beacon, which detected as little as 2% of genomic DNA from the drug-resistant cells (H1975), was much higher than direct sequencing. Furthermore, in situ fluorescence imaging with the molecular beacon gave rise to a distinguishable signal for the T790M mutation in drug-resistant cells. The molecular beacon-based approach enabled rapid and sensitive detection of the EGFR mutation (T790M) in NSCLC with in situ fluorescence imaging, which can be directed to determine various treatment options in patients with cancer.
机译：表皮生长因子受体（EGFR）中的二级突变（T790M）是对用于治疗非小细胞肺癌（NSCLC）的EGFR抑制剂获得性耐药的标志。因此，鉴定T790M突变对于指导治疗决策至关重要。鉴于DNA测序方法既耗时又不灵敏，我们开发并研究了使用分子信标检测EGFR中T790M突变的可行性。设计了与第二EGFR突变（T790M）区域互补的分子信标，并用于携带药物敏感和耐药EGFR突变的NSCLC样品。为了进行快速，简单的测定，我们尝试将分子信标与实时PCR和原位荧光成像一起使用。对来自两名具有耐药性的患者的样品测试了设计的分子信标特异性检测EGFR T790M突变的能力，并与常规DNA测序方法进行了比较。分子信标成功地在具有耐药性的患者样品中检测到T790M突变。分子信标的灵敏度比直接测序要高得多，该分子信标从耐药细胞中检出的基因组DNA仅为2％（H1975）。此外，用分子信标进行的原位荧光成像在耐药细胞中产生了T790M突变的明显信号。基于分子信标的方法可通过原位荧光成像快速，灵敏地检测NSCLC中的EGFR突变（T790M），该方法可用于确定癌症患者的各种治疗选择。

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