Communications Biology (May 2024)

Synthetic phage-based approach for sensitive and specific detection of Escherichia coli O157

  • Azumi Tamura,
  • Aa Haeruman Azam,
  • Tomohiro Nakamura,
  • Kenichi Lee,
  • Sunao Iyoda,
  • Kohei Kondo,
  • Shinjiro Ojima,
  • Kotaro Chihara,
  • Wakana Yamashita,
  • Longzhu Cui,
  • Yukihiro Akeda,
  • Koichi Watashi,
  • Yoshimasa Takahashi,
  • Hiroshi Yotsuyanagi,
  • Kotaro Kiga

DOI
https://doi.org/10.1038/s42003-024-06247-w
Journal volume & issue
Vol. 7, no. 1
pp. 1 – 13

Abstract

Read online

Abstract Escherichia coli O157 can cause foodborne outbreaks, with infection leading to severe disease such as hemolytic-uremic syndrome. Although phage-based detection methods for E. coli O157 are being explored, research on their specificity with clinical isolates is lacking. Here, we describe an in vitro assembly-based synthesis of vB_Eco4M-7, an O157 antigen-specific phage with a 68-kb genome, and its use as a proof of concept for E. coli O157 detection. Linking the detection tag to the C-terminus of the tail fiber protein, gp27 produces the greatest detection sensitivity of the 20 insertions sites tested. The constructed phage detects all 53 diverse clinical isolates of E. coli O157, clearly distinguishing them from 35 clinical isolates of non-O157 Shiga toxin-producing E. coli. Our efficient phage synthesis methods can be applied to other pathogenic bacteria for a variety of applications, including phage-based detection and phage therapy.