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Enzyme-free ultrasensitive fluorescence detection of epithelial cell adhesion molecules based on a toehold-aided DNA recycling amplification strategy†
RSC Advances ( IF 3.9 ) Pub Date : 2018-04-19 00:00:00 , DOI: 10.1039/c8ra01362d Jishun Chen 1 , Bing Shang 1 , Hua Zhang 1 , Zhengpeng Zhu 1 , Long Chen 1 , Hongmei Wang 1 , Fengying Ran 1 , Qinhua Chen 1 , Jun Chen 1
RSC Advances ( IF 3.9 ) Pub Date : 2018-04-19 00:00:00 , DOI: 10.1039/c8ra01362d Jishun Chen 1 , Bing Shang 1 , Hua Zhang 1 , Zhengpeng Zhu 1 , Long Chen 1 , Hongmei Wang 1 , Fengying Ran 1 , Qinhua Chen 1 , Jun Chen 1
Affiliation
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Epithelial cell adhesion molecules (EpCAMs) play a significant role in tumorigenesis and tumor development. EpCAMs are considered to be tumor signaling molecules for cancer diagnosis, prognosis and therapy. Herein, an enzyme-free and highly sensitive fluorescent biosensor, with a combined aptamer-based EpCAM recognition and toehold-aided DNA recycling amplification strategy, was developed for sensitive and specific fluorescence detection of EpCAMs. Due to highly specific binding between EpCAMs and corresponding aptamers, strand a, which is released from the complex of aptamer/strand a in the presence of EpCAMs which is bound to the corresponding aptamer, triggered the toehold-mediated strand displacement process. An amplified fluorescent signal was achieved by recycling strand a for ultrasensitive EpCAM detection with a detection limit as low as 0.1 ng mL−1, which was comparable or superior to that of reported immunoassays and biosensor strategies. In addition, high selectivity towards EpCAMs was exhibited when other proteins were selected as control proteins. Finally, this strategy was successfully used for the ultrasensitive fluorescence detection of EpCAMs in human serum samples with satisfactory results. Importantly, the present strategy may be also expanded for the detection of other targets using the corresponding aptamers.
中文翻译：
基于立足点辅助 DNA 回收扩增策略的上皮细胞粘附分子的无酶超灵敏荧光检测†
上皮细胞粘附分子（EpCAM）在肿瘤发生和发展中发挥着重要作用。 EpCAM 被认为是用于癌症诊断、预后和治疗的肿瘤信号分子。在此，开发了一种无酶且高灵敏度的荧光生物传感器，结合基于适体的 EpCAM 识别和立足点辅助 DNA 回收扩增策略，用于 EpCAM 的灵敏和特异性荧光检测。由于EpCAM与相应适体之间的高度特异性结合，在与相应适体结合的EpCAM存在的情况下，从适体/a链复合物中释放的a链触发了立足点介导的链置换过程。通过回收链a进行超灵敏EpCAM检测，获得了放大的荧光信号，检测限低至0.1 ng mL -1 ，这与报道的免疫测定和生物传感器策略相当或优于。此外，当选择其他蛋白质作为对照蛋白质时，表现出对 EpCAM 的高选择性。最后，该策略成功用于人血清样本中EpCAMs的超灵敏荧光检测，取得了满意的结果。重要的是，本策略还可以扩展到使用相应适体检测其他目标。
更新日期：2018-04-19
中文翻译：
基于立足点辅助 DNA 回收扩增策略的上皮细胞粘附分子的无酶超灵敏荧光检测†
上皮细胞粘附分子（EpCAM）在肿瘤发生和发展中发挥着重要作用。 EpCAM 被认为是用于癌症诊断、预后和治疗的肿瘤信号分子。在此，开发了一种无酶且高灵敏度的荧光生物传感器，结合基于适体的 EpCAM 识别和立足点辅助 DNA 回收扩增策略，用于 EpCAM 的灵敏和特异性荧光检测。由于EpCAM与相应适体之间的高度特异性结合，在与相应适体结合的EpCAM存在的情况下，从适体/a链复合物中释放的a链触发了立足点介导的链置换过程。通过回收链a进行超灵敏EpCAM检测，获得了放大的荧光信号，检测限低至0.1 ng mL -1 ，这与报道的免疫测定和生物传感器策略相当或优于。此外，当选择其他蛋白质作为对照蛋白质时，表现出对 EpCAM 的高选择性。最后，该策略成功用于人血清样本中EpCAMs的超灵敏荧光检测，取得了满意的结果。重要的是，本策略还可以扩展到使用相应适体检测其他目标。
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