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Dual-mode Au nanoprobe based on surface enhancement Raman scattering and colorimetry for sensitive determination of telomerase activity both in cell extracts and the urine of patients
摘要： Telomerase is a valuable biomarker, which is highly correlated to cancer diseases. However, single-mode probe for telomerase detection cannot satisfy the challenge of detection of telomerase activity rapidly, simply with high selectivity, sensitivity and accuracy both in preliminary diagnosis and point of care (POC) testing. Therefore, there is an urgent need to develop a robust approach with controllable assembly and high accuracy to consider both the simplification of preliminary diagnosis and POC testing and the quantification requirement for early clinical diagnosis and treatment. Herein, a novel dual-mode Au NPs probe was developed for determination of telomerase activity with controllable assembly and aggregation statement based on surface enhancement Raman scattering (SERS) and colorimetry. In this strategy, an Au dimer-based probe with high uniformity was assembled successfully, telomerase activity was reflected according to the color variations of solution and the Raman intensity of Raman reporter. Taking advantage of the uniformity of Au dimers and the combination of colorimetry and SERS techniques, our strategy determined the telomerase activity with high accuracy, sensitivity and wide range. The established probe possessed of high selectivity, sensitivity and accuracy, which was approved as a reliable, intuitional and convenient approach for detecting telomerase activity.
关键词： colorimetry,bladder cancer,surface enhanced Raman scattering,Au nanoparticles,telomerase activity
更新于2025-09-23 15:21:21
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A Conformational Switch-based Fluorescent Biosensor for Homogeneous Detection of Telomerase Activity
摘要： As a universal tumor biomarker, research on the activity and inhibition of telomerase is of great importance for cancer diagnosis and therapy. Herein, we demonstrate the conformational switch-based fluorescence detection of telomerase activity using a redesigned RNA aptamer Spinach. Briefly, the original Spinach aptamer was extended at its 5’ end and folded into an inactive conformation, where association with the small molecule fluorophore, 5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) was prevented. Only in the presence of telomerase, (TTAGGG)n repeats were added to the 3’ end of the telomerase substrate primer, and the elongation products hybridized with inactive Spinach molecules, triggering its conformational switch and refolding it into the active, DFHBI-binding conformation. Moreover, the fluorescence signal was further amplified through a target recycling circuit, where Ribonuclease H (RNase H) specifically hydrolyzed the phosphodiester bonds of RNA in the DNA-RNA hybrid. The released telomere products could then hybridize to new inactive Spinach molecules and initiate multiple amplification cycles. The proposed fluorescent biosensor presented great performance for telomerase activity detection from 100 to 5 × 104 Hela cells with a detection limit of 100 cells. Besides, this new assay offers a good biosensing platform for differentiation of cancer cell lines from normal cell line and evaluation the inhibition efficiency of telomere-binding ligand, which is of great importance for telomerase-related cancer diagnosis and therapy.
关键词： Telomerase activity,Spinach,Ribonuclease H,Inhibitor,Fluorescent biosensors
更新于2025-09-19 17:15:36
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Facile Synthesis of Fluorescent Tungsten Oxide Quantum Dots for Telomerase Detection Based on the Inner Filter Effect
摘要： The traditional detection of telomerase activity is mainly based on the polymerase chain reaction (PCR), which has the disadvantages of being time-consuming and susceptible to interference, and we here propose a facile method to fabricate fluorescent tungsten oxide quantum dots (WOx QDs) and employ them for telomerase activity sensing. It is found that the fluorescence of WOx QDs can be significantly quenched by hemin based on the inner filter effect (IFE). However, in the presence of telomerase, the primer-DNA can be extended to generate repeating unit of TTAGGG to form G-quadruplex, and thus hemin can be encapsulated to reduce its absorbance, resulting in decreased IFE and efficient fluorescence recovery of WOx QDs. Based on the fluorescence changes of IFE between hemin and WOx QDs, the telomerase activity with the range of 50-30000 HeLa cells can be detected and the lowest detection amount can reach 17 cells. The method exhibits good versatility that can be also applied to the telomerase detection from A549 and L929 cells. In addition, because of the good biocompatibility of the sensor, it can be used for real-time monitoring of telomerase activity in living cells, showing great potential in tumor diagnosis and inhibitor drug screening.
关键词： inner filter effect,G-quadruplex,hemin,telomerase activity detection,fluorescent tungsten oxide quantum dots
更新于2025-09-19 17:13:59
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Cascaded Electrochemiluminescence Signal Amplifier for the Detection of Telomerase Activity from Tumor Cells and Tissues
摘要： Telomerase is closely linked to the physiological transformation of tumor cells and is commonly overexpressed in most types of tumor cells. Therefore, telomerase has become a potential biomarker for the process of tumorigenesis, progression, prognosis and metastasis. Thus, it is important to develop a simple, accurate and reliable method for detecting telomerase activity. As a high signal-to-noise ratio mode, electrochemiluminescence (ECL) has been widely applied in the field of biomedical analysis. Here, our objective was to construct an improved ECL signal amplifier for the detection of telomerase activity. Methods: A cascaded ECL signal amplifier was constructed to detect telomerase activity with high selectivity via controllable construction of a lysine-based dendric Ru(bpy)3 2+ polymer (DRP). The sensitivity, specificity and performance index were simultaneously evaluated by standard substance and cell and tissue samples. Results: With this cascaded ECL signal amplifier, high sensitivities of 100, 50, and 100 cells for three tumor cell lines (A549, MCF7 and HepG2 cell lines) were simultaneously achieved, and desirable specificity was also obtained. Furthermore, the excellent performance of this platform was also demonstrated in the detection of telomerase in tumor cells and tissues. Conclusion: This cascaded ECL signal amplifier has the potential to be a technological innovation in the field of telomerase activity detection.
关键词： telomerase activity,signal amplifier,tumor cell,tumor tissue
更新于2025-09-09 09:28:46