Polyadenine-mediated Immobilization of Aptamers on a Gold Substrate for the Direct Detection of Bacterial Pathogens

Anal Sci. 2019 Sep 10;35(9):967-972. doi: 10.2116/analsci.19P110. Epub 2019 May 10.

Abstract

Nucleic acid aptamers have been widely used as synthetic probes for bioanalytical applications. Herein, we carried out a detailed study on the immobilization of a series of aptamers ranging from 37 to 88 bases, which are specific to either Escherichia coli (E. coli) or Staphylococcus aureus (S. aureus), on a planar gold substrate via a polyadenine-mediated immobilization method. The resultant surfaces were characterized by both surface plasmon resonance spectroscopy (SPR) and X-ray photoelectron spectroscopy. The results clearly show that the aptamer solution at a lower ionic strength gives rise to a higher lateral density of the aptamer when compared to that at a higher ionic strength. The SPR aptasensors are then employed for detecting their corresponding bacteria (i.e., E. coli and S. aureus, respectively). The data indicate that the SPR aptasensor with a higher density of aptamer exhibits a better capture of target bacteria.

Keywords: Escherichia coli; Staphylococcus aureus; Surface plasmon resonance spectroscopy; aptamer; polyadenine.

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / metabolism
  • Base Sequence
  • Bromides / chemistry
  • Escherichia coli / isolation & purification*
  • Escherichia coli / metabolism
  • Gold / chemistry*
  • Osmolar Concentration
  • Poly A / chemistry*
  • Sodium Compounds / chemistry
  • Staphylococcus aureus / isolation & purification*
  • Staphylococcus aureus / metabolism
  • Surface Plasmon Resonance / methods*

Substances

  • Aptamers, Nucleotide
  • Bromides
  • Sodium Compounds
  • polyadenine
  • Poly A
  • Gold
  • sodium bromide