• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>RSC Advances >Highly sensitive and specific screening of EGFR mutation using a PNA microarray-based fluorometric assay based on rolling circle amplification and graphene oxide
【24h】

Highly sensitive and specific screening of EGFR mutation using a PNA microarray-based fluorometric assay based on rolling circle amplification and graphene oxide

机译：基于滚动圆扩增和石墨烯基，使用基于PNA微阵列的荧光测定法的高敏感和特异性筛选EGFR突变

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Screening epidermal growth factor receptor (EGFR) mutations, especially deletions, is essential for diagnosis of non-small cell lung cancer (NSCLC) and also critical to inform treatment decisions for NSCLC patients. Here, we demonstrated a facile peptide nucleic acid (PNA) microarray-based fluorometric method for sensitive and specific detection of EGFR mutation, using rolling circle amplification (RCA), graphene oxide (GO), and a fluorescently-labeled detection probe (F-DP). First, the EGFR gene sequence was efficiently captured by the label-free PNA probe which was attached on the surface of a 96-well plate. And then, the EGFR mutation sequence was specifically amplified by RCA using the circular DNA, which was formed by the ligation of the padlock probe when hybridizing with the EGFR mutation, as a template. The single-stranded RCA product (RCAP) was then sensitively detected with the F-DP and GO system. This method has a detection limit of 0.3 pM for EGFR mutation and a high discrimination capability to target EGFR mutation against EGFR wildtype. The use of a PNA microarray and a fluorescence quenching platform make this system quite suitable for high-throughput analysis of EGFR mutations in resource-limited settings without the need of costly and cumbersome equipment. Furthermore, this detection system provides a novel way for the diagnosis of other diseases that are caused by gene deletion mutations.
机译：筛选表皮生长因子受体（EGFR）突变，尤其是缺失，对于诊断非小细胞肺癌（NSCLC），对NSCLC患者的治疗决策也至关重要。在这里，我们证明了一种容易肽核酸（PNA）微阵列的荧光测定方法，用于使用滚动圆扩增（RCA），石墨烯（GO）和荧光标记的检测探针（F- DP）。首先，通过附着在96孔板的表面上，通过无标记的PNA探针有效地捕获EGFR基因序列。然后，使用圆形DNA通过RCA特异性扩增EGFR突变序列，该圆形DNA通过与EGFR突变杂交时通过连接挂锁探针而形成，作为模板。然后用F-DP和GO系统敏感地检测单链RCA产品（RCAP）。该方法具有0.3μm的检出限为EGFR突变和对EGFR野生型靶向EGFR突变的高鉴别能力。使用PNA微阵列和荧光猝灭平台使该系统非常适合资源限制环境中EGFR突变的高通量分析，而无需昂贵和繁琐的设备。此外，该检测系统为诊断由基因缺失突变引起的其他疾病提供了一种新的方法。

著录项

  • 来源
    《RSC Advances》 |2019年第66期|共11页
  • 作者

  • 作者单位
  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
  • 关键词

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号