Strand displacement amplification-coupled dynamic light scattering method to detect urinary telomerase for non-invasive detection of bladder cancer

Biosens Bioelectron. 2019 Apr 15:131:143-148. doi: 10.1016/j.bios.2019.02.014. Epub 2019 Feb 18.

Abstract

Despite huge successes achieved by strand displacement amplification (SDA) and gold nanoparticles (AuNPs) in biomolecules sensing, the strategy of combination of SDA and AuNPs-based dynamic light scattering (DLS) for a biomolecule sensing is unexplored. Here we developed a non-invasive, SDA-based DLS method for the diagnosis of bladder cancer by detecting telomerase activity in human urine. In the presence of telomerase, the telomerase substrate (TS) primer was elongated with repeating sequences of (TTAGGG)n, and the resulting product triggers SDA between the hairpin deoxyribonucleic acid (DNA) and the Primer. The SDA product can be recognized by the oligonucleotide-modified AuNPs probes, resulting in DLS measurable AuNPs aggregation. The assay displayed a detection limit of 3 MCF-7 cells with a signal-to-noise ratio of 3 in a dynamic range of 5-1000 cells. The method was simple, reliable and has been successfully applied in the detection of telomerase in urine with good accuracy, selectivity and reproducibility. Moreover, only urine samples from bladder cancer patients induced a significant change in the average hydrodynamic diameter, indicating practical applicability of the method for the non-invasive diagnosis of bladder cancer.

Keywords: Bladder cancer; Detection; Dynamic light scattering; Gold nanoparticles; Strand displacement amplification; Telomerase activity.

MeSH terms

  • Biosensing Techniques*
  • Dynamic Light Scattering
  • HeLa Cells
  • Humans
  • Limit of Detection
  • Metal Nanoparticles / chemistry
  • Oligonucleotides / chemistry
  • Telomerase / chemistry
  • Telomerase / isolation & purification*
  • Telomerase / urine
  • Urinary Bladder Neoplasms / pathology
  • Urinary Bladder Neoplasms / urine*

Substances

  • Oligonucleotides
  • Telomerase