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滚环扩增结合核酸外切酶III辅助显色反应用于灵敏的端粒酶活性分析

Rolling Circle Amplification Integrating with Exonuclease-III-Assisted Color Reaction for Sensitive Telomerase Activity Analysis.

作者信息

Liu Xiaoya, Zhao Xianxian, Zhang Jie, Wang Yihan, Ye Xiaoping

机构信息

Department of Oncology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.

Central Laboratory, Chongqing University FuLing Hospital, Chongqing 408099, China.

出版信息

ACS Omega. 2024 Dec 4;9(50):49081-49087. doi: 10.1021/acsomega.4c03839. eCollection 2024 Dec 17.

Abstract

Telomerase activation can lead to the escape from cell senescence and immortalization, playing a crucial role in the growth and proliferation of cancer cells. Therefore, the detection of telomerase activity is essential for cancer diagnosis and treatment. Herein, we develop a novel ultrasensitive and visually detectable platform. By incorporation of exonuclease-III (Exo-III), this platform achieves dual signal amplification of rolling circle amplification products. Additionally, the colorimetric analysis of 3,3',5,5'-tetramethylbiphenyl (TMB) chromogenic reaction system provides this approach with unique advantages such as simplicity, speediness, and sensitivity. The detection platform exhibits high sensitivity and specificity in actual sample testing, which aligns closely with results obtained using commercial kits. Moreover, it offers ease-of-use through visual determination by the naked eyes. This finding indicates that our proposed sensing method performs satisfactorily in detecting telomerase in real biological samples. Henceforth, we believe that this sensing platform holds great potential for clinical diagnosis and anticancer drug development.

摘要

端粒酶激活可导致细胞逃脱衰老并实现永生化,在癌细胞的生长和增殖中起关键作用。因此,端粒酶活性的检测对于癌症诊断和治疗至关重要。在此,我们开发了一种新型的超灵敏且可视觉检测的平台。通过掺入核酸外切酶III(Exo-III),该平台实现了滚环扩增产物的双信号放大。此外,3,3',5,5'-四甲基联苯(TMB)显色反应体系的比色分析为该方法提供了诸如简单、快速和灵敏等独特优势。该检测平台在实际样品测试中表现出高灵敏度和特异性,与使用商业试剂盒获得的结果密切相符。此外,它通过肉眼视觉判定实现了易用性。这一发现表明我们提出的传感方法在检测实际生物样品中的端粒酶方面表现令人满意。从今往后,我们相信这个传感平台在临床诊断和抗癌药物开发方面具有巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4f12/11656203/6aefa90f797b/ao4c03839_0001.jpg

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