4.5 Article

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

Journal

ANALYTICAL BIOCHEMISTRY
Volume 551, Issue -, Pages 37-42

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2018.04.015

Keywords

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

Funding

  1. Key Research and Development Program of Jiangsu Province [BE2016306]
  2. National Science and Technology Support Program of China [2015BAD17B02]
  3. Sichuan Science and Technology Program [2018JY0194, 2018SZ0340]
  4. Key scientific research fund of Xihua University [Z1620514]
  5. Key Laboratory Open Research Fund of Grain and Oil Processing and Food Safety of Xihua University [szjj2016-022]

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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 (K-d) 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.

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