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Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR
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DNA aptamer identification and characterization for E. coli O157 detection using cell based SELEX method.

Masoum Amraee1, Mana Oloomi1, Afsaneh Yavari1

  • 1Department of Molecular Biology, Pasteur Institute of Iran, Pasteur Ave., Tehran 13164, Iran.

Analytical Biochemistry
|August 19, 2017
PubMed
Summary

Researchers developed a specific DNA aptamer to rapidly detect Escherichia coli (E. coli) O157:H7, a dangerous foodborne pathogen. This aptamer offers a promising tool for identifying E. coli O157:H7 in foodborne disease diagnostics.

Keywords:
Cell SELEXDNA aptamerDissociation constant

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Food Safety

Background:

  • Escherichia coli (E. coli) O157:H7 is a significant foodborne pathogen causing human illness.
  • Rapid identification of E. coli O157:H7 is crucial to mitigate its toxic effects.

Purpose of the Study:

  • To develop a specific DNA aptamer for the detection and capture of E. coli O157:H7.
  • To evaluate the efficiency and specificity of the selected aptamer.

Main Methods:

  • Systematic evolution of ligands by exponential enrichment using whole cells (Cell-SELEX) was employed.
  • Nine rounds of Cell-SELEX were performed using E. coli O157 as the target and other E. coli strains and related bacteria as counter-targets.
  • Candidate aptamer sequences were selected and their specificity, dissociation constant, and secondary structure were analyzed.

Main Results:

  • A specific DNA aptamer was successfully selected for E. coli O157 detection and capture.
  • The selected aptamer exhibited a dissociation constant of 107.6 ± 67.8 pM.
  • Cross-reactivity assays confirmed the aptamer's specificity for E. coli O157.

Conclusions:

  • The developed DNA aptamer is highly efficient and specific for detecting E. coli O157:H7.
  • This aptamer holds potential as a diagnostic ligand for identifying foodborne diseases caused by E. coli O157:H7.