Design and development of PCR-free highly sensitive electrochemical assay for detection of telomerase activity using Nano-based (liposomal) signal amplification platform

Biosens Bioelectron. 2016 Jun 15:80:426-432. doi: 10.1016/j.bios.2016.01.090. Epub 2016 Feb 1.

Abstract

Telomerase, which has been detected in almost all kinds of cancer tissues, is considered as an important tumor marker for early cancer diagnostics. In the present study, an electrochemical method based on liposomal signal amplification platform is proposed for simple, PCR-free, and highly sensitive detection of human telomerase activity, extracted from A549 cells. In this strategy, telomerase reaction products, which immobilized on streptavidin-coated microplate, hybridized with biotinylated capture probes. Then, dopamine-loaded biotinylated liposomes are attached through streptavidin to biotinylated capture probes. Finally, liposomes are ruptured by methanol and the released-dopamine is subsequently measured using differential pulse voltammetry technique by multi-walled carbon nanotubes modified glassy carbon electrode. Using this strategy, the telomerase activity extracted from 10 cultured cancer cells could be detected. Therefore, this approach affords high sensitivity for telomerase activity detection and it can be regarded as an alternative to telomeric repeat amplification protocol assay, having the advantages of simplicity and less assay time.

Keywords: Carbon nanotubes; Differential pulse voltammetry; Dopamine; Liposome; Signal amplification; Telomerase activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / chemistry
  • Biomarkers, Tumor / isolation & purification*
  • Biosensing Techniques*
  • HeLa Cells
  • Humans
  • Liposomes / chemistry
  • Nanotubes, Carbon / chemistry
  • Neoplasms / diagnosis*
  • Neoplasms / genetics
  • Telomerase / chemistry
  • Telomerase / genetics
  • Telomerase / isolation & purification*

Substances

  • Biomarkers, Tumor
  • Liposomes
  • Nanotubes, Carbon
  • Telomerase