A novel DNA biosensor integrated with Polypyrrole/streptavidin and Au-PAMAM-CP bionanocomposite probes to detect the rs4839469 locus of the vangl1 gene for dysontogenesis prediction

Biosens Bioelectron. 2016 Jun 15:80:674-681. doi: 10.1016/j.bios.2016.02.025. Epub 2016 Feb 10.

Abstract

The single nucleotide polymorphism (SNP) of the vangl1 gene is highly correlated with Neural Tube Defects (NTDs), a group of severe congenital malformations. It is hindered by the lack of a quantitative detection method. We first propose the use of a DNA biosensor to detect the missense single nucleotide polymorphism (rs4839469 c.346G>A p.Ala116Thr) of the vangl1 gene in this work. Polypyrrole (PPy) and streptavidin were integrated to modify a gold electrode. We took advantage of the PPy's good biocompatibility and excellent conductivity. To further accelerate the electron transfer process at the electrode surface, polyamidoamine dendrimer-encapsulated gold nanoparticles (Au-PAMAM) were used, because Au-PAMAM possess a large number of amino groups to load capture probes (CP). Using the biotin-streptavidin system, the Au-PAMAM-CP bionanocomposite probe, which can detect the target DNA, was conjugated to the electrode surface. Under optimal conditions, the DNA biosensor exhibited a wide linear range of 0.1-100 nM with a low detection limit of 0.033 nM (S/N=3). The results suggest that this approach has the potential to be used in clinical research.

Keywords: Au-PAMAM; Capture probe; DNA biosensor; PPy/ streptavidin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques*
  • Carrier Proteins / genetics
  • Carrier Proteins / isolation & purification*
  • DNA / genetics
  • DNA / isolation & purification*
  • Gold / chemistry
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / isolation & purification*
  • Nanocomposites / chemistry
  • Neural Tube Defects / diagnosis*
  • Neural Tube Defects / genetics
  • Polymers / chemistry
  • Polymorphism, Single Nucleotide / genetics
  • Pyrroles / chemistry
  • Streptavidin / chemistry

Substances

  • Carrier Proteins
  • Membrane Proteins
  • Polymers
  • Pyrroles
  • VANGL1 protein, human
  • polypyrrole
  • Gold
  • DNA
  • Streptavidin