The Use of a Novel NanoLuc -Based Reporter Phage for the Detection of Escherichia coli O157:H7
Rapid detection of the foodborne pathogen Escherichia coli O157:H7 is of vital importance for public health worldwide. Among detection methods, reporter phages represent unique and sensitive tools for the detection of E. coli O157:H7 from food as they are host-specific and able to differentiate live...
Gespeichert in:
Veröffentlicht in: | Scientific reports 2016-09, Vol.6 (1), p.33235, Article 33235 |
---|---|
Hauptverfasser: | , , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
Zusammenfassung: | Rapid detection of the foodborne pathogen
Escherichia coli
O157:H7 is of vital importance for public health worldwide. Among detection methods, reporter phages represent unique and sensitive tools for the detection of
E. coli
O157:H7 from food as they are host-specific and able to differentiate live cells from dead ones. Upon infection, target bacteria become identifiable since reporter genes are expressed from the engineered phage genome. The
E. coli
O157:H7 bacteriophage ΦV10 was modified to express NanoLuc luciferase (Nluc) derived from the deep-sea shrimp
Oplophorus gracilirostris
. Once infected by the ΦV10 reporter phage,
E. coli
O157:H7 produces a strong bioluminescent signal upon addition of commercial luciferin (Nano-Glo
®
). Enrichment assays using
E. coli
O157:H7 grown in LB broth with a reporter phage concentration of 1.76 × 10
2
pfu ml
−1
are capable of detecting approximately 5 CFU in 7 hours. Comparable detection was achieved within 9 hours using 9.23 × 10
3
pfu ml
−1
of phage in selective culture enrichments of ground beef as a representative food matrix. Therefore we conclude that this NanoLuc reporter phage assay shows promise for detection of
E. coli
O157:H7 from food in a simple, fast and sensitive manner. |
---|---|
ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/srep33235 |