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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >A novel high-resolution chipCE assay for rapid detection of EGFR gene mutations and amplifications in lung cancer therapy by a combination of fragment analysis, denaturing CE and MLPA.
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A novel high-resolution chipCE assay for rapid detection of EGFR gene mutations and amplifications in lung cancer therapy by a combination of fragment analysis, denaturing CE and MLPA.

机译：通过结合片段分析，CE和MLPA变性，可在肺癌治疗中快速检测EGFR基因突变和扩增的新型高分辨率chipCE检测方法。

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摘要

There is a growing interest in evaluating molecular markers as predictors of response to new generation of targeted cancer therapies. One of such areas is biological therapy targeting epidermal growth factor receptor gene (EGFR) in lung cancer. The testing of tumor tissue is focused on specific EGFR mutations and EGFR gene amplification, since tumors exhibiting positivity of either of the two marker types are highly sensitive towards the treatment. Although traditional methods of DNA sequencing and fluorescence in situ hybridization are still in use for the detection of EGFR mutations and gene amplification, respectively, there is a need for new dedicated techniques with the primary emphasis on simplicity, sensitivity, speed and cost effectiveness. The main purpose of this work was to integrate diverse assays for both EGFR tests onto a single platform to eliminate the need for different instruments and separate processing. We demonstrate a chip capillary electrophoresis (chipCE) application for EGFR mutation detection by a combination of fragment analysis and denaturing CE along with multiplex ligation-dependent probe amplification (MLPA) for evaluation of EGFR amplification. All separations are carried out in denaturing sieving polymer on a modified Bioanalyzer 2100 chipCE instrument running at temperatures of up to 65 degrees C. The main strength of the resulting high-resolution chipCE application is in its simplicity, speed of analysis and minimal amount of sample required for complete testing of EGFR status. Such an approach could potentially fit medium throughput laboratories providing molecular pathology services for clinical oncologists with fast turnaround times and limited consumption of tissue material.
机译：评估分子标记物作为对新一代靶向癌症治疗反应的预测因子的兴趣日益浓厚。这些领域之一是针对肺癌的表皮生长因子受体基因（EGFR）的生物疗法。肿瘤组织的测试集中在特定的EGFR突变和EGFR基因扩增上，因为表现出两种标记物类型之一阳性的肿瘤对治疗高度敏感。尽管传统的DNA测序和荧光原位杂交方法仍分别用于检测EGFR突变和基因扩增，但仍需要新的专用技术，其主要重点是简便性，敏感性，速度和成本效益。这项工作的主要目的是将两种EGFR检测的多种检测方法整合到一个平台上，从而消除了对不同仪器和单独处理的需求。我们通过结合片段分析和变性CE以及多重连接依赖探针扩增（MLPA）来评估EGFR扩增，证明了芯片毛细管电泳（chipCE）在EGFR突变检测中的应用。所有分离均在经过改进的Bioanalyzer 2100 chipCE仪器上变性聚合物筛分中进行，该仪器在高达65摄氏度的温度下运行。所得高分辨率chipCE应用的主要优势在于其简便性，分析速度和最少的样品量完整测试EGFR状态所需的。这种方法可能适合中等吞吐量的实验室，为临床肿瘤学家提供分子病理学服务，具有快速的周转时间和有限的组织材料消耗。

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