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Accurate and sensitive detection of Salmonella in foods by engineered bacteriophages

Authors
  • Nguyen, Minh M.1
  • Gil, Jose2
  • Brown, Matthew3
  • Cesar Tondo, Eduardo4
  • Soraya Martins de Aquino, Nathanyelle4
  • Eisenberg, Marcia3
  • Erickson, Stephen1
  • 1 Laboratory Corporation of America Holdings, New Brighton, MN, 55112, USA , New Brighton (United States)
  • 2 Laboratory Corporation of America Holdings, Los Angeles, CA, 90062, USA , Los Angeles (United States)
  • 3 Laboratory Corporation of America Holdings, Burlington, NC, 27215, USA , Burlington (United States)
  • 4 Instituto de Ciência e Tecnologia de Alimentos, Universidade Federal do Rio Grande do Sul (ICTA/UFRGS), Porto Alegre, RS, 91501-970, Brazil , Porto Alegre (Brazil)
Type
Published Article
Journal
Scientific Reports
Publisher
Springer Nature
Publication Date
Oct 15, 2020
Volume
10
Issue
1
Identifiers
DOI: 10.1038/s41598-020-74587-8
Source
Springer Nature
License
Green

Abstract

Salmonella is a major causative agent of foodborne illness and rapid identification of this pathogen is essential to prevent disease. Currently most assays require high bacterial burdens or prolonged enrichment to achieve acceptable performance. A reduction in testing time without loss of sensitivity is critical to allow food processors to safely decrease product holding time. To meet this need, a method was developed to detect Salmonella using luciferase reporter bacteriophages. Bacteriophages were engineered to express NanoLuc, a novel optimized luciferase originating from the deep-sea shrimp Oplophorus gracilirostris. NanoLuc-expressing bacteriophages had a limit of detection of 10–100 CFU per mL in culture without enrichment. Luciferase reporters demonstrated a broad host range covering all Salmonella species with one reporter detecting 99.3% of 269 inclusivity strains. Cross-reactivity was limited and only observed with other members of the Enterobacteriaceae family. In food matrix studies, a cocktail of engineered bacteriophages accurately detected 1 CFU in either 25 g of ground turkey with a 7 h enrichment or 100 g of powdered infant formula with a 16 h enrichment. Use of the NanoLuc reporter assay described herein resulted in a considerable reduction in enrichment time without a loss of sensitivity.

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