A simple colorimetric detection of telomerase exploiting specific cleavage of exonuclease III coupled with telomeric DNA controlled aggregation of nanogold.

Abstract

Telomerase activity has piqued scientists' interest for the reason that it has the potential to be employed for early-stage cancer detection, anticancer therapy and studies related to cancer progression and metastasis. Several approaches have been developed to detect telomerase activity. However, these approaches were lengthy, challenging to quantify, of limited sensitivity and prone to polymerase chain reaction (PCR)-related artefacts. We herein developed a novel nanoplasmonic sensing platform for colorimetric detection of telomerase activity relying on the telomere elongation of telomerase at the 3' end, structure-specific cleavage activity of exonuclease III that removes mononucleotides from the 3'-hydroxyl termini of double-stranded DNA, and electrostatic interaction of elongated telomeres with plasmonic nanoparticles. Using HeLa cells as a model for colorimetric detection of telomerase activity, this biosensor could detect telomerase activity with a detection limit of ∼100 cells per reaction by visible inspection and ∼5 cells per reaction by spectroscopic measurement and analysis time within about three hours. The proposed sensing method provided a novel tool for simple, rapid, and low-cost detection of telomerase activity, eliminating the necessity for thermal cycling, primers in PCR-based assays, and amplification of telomerase extension products. It exhibits significant potential as a label-free, simple, ultrasensitive strategy for on-site detection of telomerase activity in proteomics and clinical diagnostics.