Quantitative Determination of Telomerase Activity by Combining Fluorescence Correlation Spectroscopy with Telomerase Repeat Amplification Protocol

Su, Di; Huang, Xiangyi; Dong, Chaoqing; Ren, Jicun

doi:10.1021/acs.analchem.7b04256

Cited by 22 publications

(6 citation statements)

References 45 publications

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“…A recent alternative method to determine telomerase activity in human urine samples using the hybridization chain reaction and dynamic light scattering has been developed for the detection of bladder cancer [23]. Preliminary findings indicate a high specificity in cellular models and in few healthy individuals and patients with malignancies other than bladder cancer [24][25][26], but this needs to be confirmed in large case-control studies.…”

Section: Telomerase Reactivation In Bladder Cancer and Early-generation Of Telomerase Urinary-based Biomarkersmentioning

confidence: 99%

Activating Telomerase TERT Promoter Mutations and Their Application for the Detection of Bladder Cancer

Zvereva

Pisarev

Hosen

et al. 2020

IJMS

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This review summarizes state-of-the-art knowledge in early-generation and novel urine biomarkers targeting the telomerase pathway for the detection and follow-up of bladder cancer (BC). The limitations of the assays detecting telomerase reactivation are discussed and the potential of transcription-activating mutations in the promoter of the TERT gene detected in the urine as promising simple non-invasive BC biomarkers is highlighted. Studies have shown good sensitivity and specificity of the urinary TERT promoter mutations in case-control studies and, more recently, in a pilot prospective cohort study, where the marker was detected up to 10 years prior to clinical diagnosis. However, large prospective cohort studies and intervention studies are required to fully validate their robustness and assess their clinical utility. Furthermore, it may be interesting to evaluate whether the clinical performance of urinary TERT promoter mutations could increase when combined with other simple urinary biomarkers. Finally, different approaches for assessment of TERT promoter mutations in urine samples are presented together with technical challenges, thus highlighting the need of careful technological validation and standardization of laboratory methods prior to translation into clinical practice.

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Section: Telomerase Reactivation In Bladder Cancer and Early-generation Of Telomerase Urinary-based Biomarkersmentioning

confidence: 99%

Activating Telomerase TERT Promoter Mutations and Their Application for the Detection of Bladder Cancer

Zvereva

Pisarev

Hosen

et al. 2020

IJMS

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“…More importantly, FCS is especially suitable for in situ and real-time study in live cells. 6,7 We have been engaging in FCS and its application for a long time, and we successfully applied FCS in homogeneous immunoassay, 8 cell-free DNA analysis, 9 cell apoptosis detection, 10 drug screening, 11,12 detection of enzyme activity, 13 characterization on quantum dot (QDs), 14,15 and in vivo detection on molecular dynamics 16 and molecular interactions. 17−19 The detection capability of single-molecule/particle optical detection methods largely depends on the properties of the used optical probes.…”

Section: Introductionmentioning

confidence: 99%

Resonance Light-Scattering Correlation Spectroscopy and Its Application in Analytical Chemistry for Life Science

Dong,

Ren

2023

Acc. Chem. Res.

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Conspectus Resonance light-scattering correlation spectroscopy (RLSCS) is a new single-particle detection method with its working principle being like fluorescence correlation spectroscopy (FCS). RLSCS is obtained by autocorrelation function analysis on the measured fluctuation of the resonance light scattering (RLS) intensity occurring within a subfemtoliter volume when a single nanoparticle (such as gold nanoparticles (NPs) or silver (SNPs)) freely diffuses through the volume. The RLSCS technique can detect such parameters as concentration, diffusion coefficient (translation and rotation), etc. Compared with the FCS technique, the correlated fluorescence intensity signal in RLSCS is replaced with the RLS signal of the nanoparticles, overcoming some limits of the fluorescent probes such as photobleaching under high-intensity or long-term illumination. In this Account, we showcase RLSCS methods, theoretical models at different optical configurations, and some key applications. First, the RLSCS optical detection system was constructed based on the confocal optics, its theoretical model was proposed, and the diffusion behaviors of the nanoparticles in the solution were studied including the rotational and translational diffusion. And, methods were developed to measure the concentration, size, aspect ratio, and size distribution of the NPs. Second, based on the RLSCS methods, some detection strategies were developed for homogeneous DNA detection, immunoassay, apoptosis assay, self-thermophoresis of the nanomotor, and quantitative assay in single living cells. Meanwhile, a new fluorescence/scattering cross-correlation spectroscopy (FSCCS) method was proposed for monitoring the molecule–particle interaction. This method enriched the conventional fluorescence/fluorescence cross-correlation spectroscopy (FCCS) method. Third, using the EMCCD with high sensitivity and rapid response as an optical detector, two temporospatially resolved scattering correlation spectroscopy methods and their theoretical models were developed: total internal reflection (TIR) configuration-based spatially resolved scattering correlation spectroscopy (SRSCS) and dark-field illumination-based scattering correlation spectroscopy (DFSCS). These methods extended single-spot confocal RLSCS to imaging RLSCS, which makes RLSCS have the ability for multiple channel detection with temporospatial resolution. The method was successfully used for investigating the dynamic behaviors of gold NPs in live cells and obtained its temporospatial concentration distribution and diffusion behaviors. The final section of this Account outlines future directions in the development of RLSCS.

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“…Therefore, telomerase is regarded as a cancer marker and it is necessary to develop a sensitive and reliable sensing platform for detecting telomerase activity. To date, several smart strategies have been proposed for the detection of telomerase activity . In these methods, a molecular beacon (MB)‐like probe shows its designability as a widely used tool.…”

Section: Introductionmentioning

confidence: 99%

A Fluorescence Light‐Up Silver Nanocluster Beacon Modulated by Metal Ions and Its Application in Telomerase‐Activity Detection

Peng

Ouyang

2019

Chemistry A European J

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DNA‐templated silver nanoclusters (DNA‐AgNCs) have been extensively studied in recent years. The enhancement of fluorescence emission from DNA‐AgNCs is still being explored. Herein, a study on the fluorescence enhancement of DNA‐AgNCs induced by metal ions is reported. The enhancement is greatly dependent on the primary sequence and secondary structure of the DNA strands. Thus, a label‐free AgNCs‐based molecular beacon (MB) was explored for the detection of telomerase activity. Nonfluorescent MB‐AgNCs in phosphate buffer emit dramatic red fluorescence when Mg2+ is introduced, whereas Mg2+ has a limited effect on the weak fluorescence of DNA‐AgNCs when the hairpin structure of MB is opened. Telomerase primer can be elongated by telomerase, which results in unfolding of MB in a strand‐displacement reaction. On the basis of the different brightnesses of AgNCs produced by the two DNA templates, telomerase activity can be detected. The MB‐AgNCs sensing platform provides a simple and low‐cost method to detect telomerase activity and shows great potential in the construction of cost‐effective probes for biomolecular detection.

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Quantitative Determination of Telomerase Activity by Combining Fluorescence Correlation Spectroscopy with Telomerase Repeat Amplification Protocol

Cited by 22 publications

References 45 publications

Activating Telomerase TERT Promoter Mutations and Their Application for the Detection of Bladder Cancer

Activating Telomerase TERT Promoter Mutations and Their Application for the Detection of Bladder Cancer

Resonance Light-Scattering Correlation Spectroscopy and Its Application in Analytical Chemistry for Life Science

A Fluorescence Light‐Up Silver Nanocluster Beacon Modulated by Metal Ions and Its Application in Telomerase‐Activity Detection

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