Selection and characterization, application of a DNA aptamer targeted to Streptococcus pyogenes in cooked chicken

Anal Biochem. 2018 Jun 15:551:37-42. doi: 10.1016/j.ab.2018.04.015. Epub 2018 Apr 23.

Abstract

An aptamer against Streptococcus pyogenes was selected and identified, and a fluorescent method based on the reported aptamer was established to detect S. pyogenes in the cooked chicken. Through a twelve rounds of whole-bacterium SELEX (systematic evolution of ligands by exponential enrichment) selection in vitro, a set of aptamers binding to the whole cell of S. pyogenes were generated, harvesting a low-level dissociation constant (Kd) value of 44 ± 5 nmol L-1 of aptamer S-12. Aptamer-based quantification of S. pyogenes in the cooked chicken sample was implemented in a fluorescence resonance energy transfer-based assay by using graphene oxide, resulting in a limit of detection of 70 cfu mL-1. The selected aptamer showed affinity and selectivity recognizing S. pyogenes; besides, more biosensors based on the selected aptamer as a molecular recognition element could be developed in the innovative determinations of S. pyogenes.

Keywords: Aptamer; Cooked chicken; Graphene oxide; Streptococcus pyogenes; Whole-bacterium SELEX.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aptamers, Nucleotide / chemistry
  • Aptamers, Nucleotide / genetics*
  • Base Sequence
  • Chickens / microbiology*
  • Cooking
  • DNA, Bacterial / genetics
  • Fluorescence Resonance Energy Transfer
  • Food Microbiology* / methods
  • Graphite
  • Nucleic Acid Conformation
  • SELEX Aptamer Technique / methods
  • Streptococcus pyogenes / genetics*
  • Streptococcus pyogenes / isolation & purification
  • Streptococcus pyogenes / pathogenicity

Substances

  • Aptamers, Nucleotide
  • DNA, Bacterial
  • graphene oxide
  • Graphite