Ratiometric Fluorescent Bioprobe for Highly Reproducible Detection of Telomerase in Bloody Urines of Bladder Cancer Patients

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• 作者：Yuan Zhuang ; Qi Xu ; Fujian Huang ; Pengcheng Gao ; Zujin Zhao ; Xiaoding Lou ; Fan Xia
• 刊名：ACS Sensor
• 出版年：2016
• 出版时间：May 27, 2016
• 年：2016
• 卷：1
• 期：5
• 页码：572-578
• 全文大小：593K
• ISSN：2379-3694

文摘

Fluorescent bioprobes, as one of the most important tools, hold high promise for real-time analytical sensing of biological molecules and processes in live cells and organisms. Although several excellent bioprobes employ turn-on fluorescence intensity, such a sole responsive signal is readily perturbed by various experimental conditions. Herein, to improve the reproducibility and robustness, a ratiometric fluorescent bioprobe for telomerase activity detection has been developed. The ratiometric fluorescent bioprobe is designed on the use of two fluorescence dyes in parallel. One is red emissive aggregation-caused quenching (ACQ) dye, Cy5, as a control, molecularly labeled on the 5′-end of telomerase substrate oligonucleotides (TS primer). The other is water-soluble aggregation-induced emission (AIE) dye, Silole-R, as reporter of telomerase activity, nonemissive in buffer. In the presence of telomerase, the blue emission is enhanced by the added negatively charged sites for Silole-R to bind and aggregate, while the red emission is almost unchanged as stable internal reference. With the addition of incremental amounts of telomerase, the ratiometric emission intensity ratios (I₄₇₈/I₆₆₅) of this bioprobe gradually increase. Furthermore, the distinguishing of telomerase extracts from 20 bladder cancer bloody and 10 normal urine specimens confirms the practicality of this bioprobe. In contrast to previous turn-on bioprobes, these advanced experiments obtain higher reproducibility and positive result rate (100%) toward bladder cancer bloody urine specimens.