Sensitive naked-eye detection of telomerase activity based on exponential amplification reaction and lateral flow assay
Sensitive naked-eye detection of telomerase activity based on exponential amplification reaction and lateral flow assay
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DOI：
10.1007/s00216-022-04179-0
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发表时间：
2022-06-17
影响因子：
4.3
通讯作者：
Li, Xin-Min
中科院分区：
文献类型：
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作者：
Cheng, Xue-Ru;Wang, Fei;Li, Xin-Min
端粒酶是一种很有前途的癌症诊断和预后生物标志物。灵敏、简便、可靠的端粒酶活性检测对肿瘤诊断具有重要意义。在此，我们建立了一种结合指数扩增反应(EXPAR)和侧向流动分析的超灵敏的端粒酶活性可视化检测方法，我们称之为侧向流动读出-EXPAR(LFR-EXPAR)。在LFR-EXPAR检测中，端粒酶延长产物启动指数扩增反应，产生的触发器与报告分子杂交形成裂口酶的识别位点，裂口酶切割报告链。报告器的退化可以用通用的侧向流量尺检测，并用肉眼读出。在进行了一系列概念验证研究后，LFR-EXPAR分析被发现获得了与TRAP(端粒重复序列扩增协议)相当的灵敏度。LFR-EXPAR法可以实现对端粒酶的超灵敏和定点检测，而不需要专门的仪器，为癌症的早期诊断带来了巨大的希望。
Telomerase is a promising diagnostic and prognostic biomarker for cancers. Sensitive, simple, and reliable telomerase activity detection is vital for cancer diagnosis. Herein, we developed an ultrasensitive visualized assay for telomerase activity that combined the exponential amplification reaction (EXPAR) and lateral flow assay for easy and quick signal readout, which we termed as a lateral flow readout-EXPAR (LFR-EXPAR) assay. In the LFR-EXPAR assay, telomerase elongation products initiate the exponential amplification reaction, the generated trigger hybridizes with the reporter to form the recognition site of the nicking enzyme, and the nicking enzyme cuts the reporter strand. The degradation of the reporter can be detected with a universal lateral flow dipstick and read out with the naked eye. After conducting a series of proof-of-concept investigations, the LFR-EXPAR assay was found to achieve a sensitivity comparable to that of a TRAP (telomere repeat amplification protocol) assay. The LFR-EXPAR assay can be used to realize ultrasensitive and point-of-care detection of telomerase without requiring specialized instruments, holding great promise for early cancer diagnosis.