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 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.
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Dandan
D
Zhang
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Claudia P
CP
Coronel-Aguilera
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Patricia L
PL
Romero
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Lynda
L
Perry
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Udit
U
Minocha
Department of Biological Science, Purdue University, West Lafayette, IN 47907, USA.
Carla
C
Rosenfield
Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, PA 19038, USA.
Andrew G
AG
Gehring
Molecular Characterization of Foodborne Pathogens Research Unit, Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, PA 19038, USA.
George C
GC
Paoli
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
Arun K
AK
Bhunia
Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.|Department of Biological Science, Purdue University, West Lafayette, IN 47907, USA.
Bruce
B
Applegate
D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D002417:Cattle; D015169:Colony Count, Microbial; D019453:Escherichia coli O157 / Q000302:isolation & purification* / Q000472:pathogenicity; D005506:Food Contamination / Q000032:analysis; D005516:Food Microbiology / Q000379:methods*; D008156:Luciferases / Q000737:chemistry* / Q000235:genetics; D008460:Meat / Q000382:microbiology
D008156:Luciferases