Article ; Online: Sensitive detection of viable Cronobacter sakazakii by bioluminescent reporter phage emitting stable signals with truncated holin.
Food research international (Ottawa, Ont.)
2023 Volume 174, Issue Pt 2, Page(s) 113665
Abstract: As outbreaks of foodborne illness caused by the opportunistic pathogen Cronobacter sakazakii (Cs) continue to occur, particularly in infants consuming powdered infant formula (PIF), the need for sensitive, rapid, and easy-to-use detection of Cs from food ...
| Abstract | As outbreaks of foodborne illness caused by the opportunistic pathogen Cronobacter sakazakii (Cs) continue to occur, particularly in infants consuming powdered infant formula (PIF), the need for sensitive, rapid, and easy-to-use detection of Cs from food and food processing environments is increasing. Here, we developed bioluminescent reporter bacteriophages for viable Cs-specific, substrate-free, rapid detection by introducing luciferase and its corresponding substrate-providing enzyme complex into the virulent phage ΦC01. Although the reporter phage ΦC01_lux, constructed by replacing non-essential genes for phage infectivity with a luxCDABE reporter operon, produced bioluminescence upon Cs infection, the emitted signal was quickly decayed due to the superior bacteriolytic activity of ΦC01. By truncating the membrane pore-forming protein holin and thus limiting its function, the bacterial lysis was delayed and the resultant engineered reporter phage ΦC01_lux_Δhol could produce a more stable and reliable bioluminescent signal. Accordingly, ΦC01_lux_Δhol was able to detect at least an average of 2 CFU/ml of Cs artificially contaminated PIF and Sunsik and food contact surface models within a total of 7 h of assays, including 5 h of pre-enrichment for Cs amplification. The sensitive, easy-to-use, and specific detection of live Cs with the developed reporter phage could be applied as a novel complementary tool for monitoring Cs in food and food-related environments for food safety and public health. |
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| MeSH term(s) | Cronobacter sakazakii/genetics ; Cronobacter sakazakii/isolation & purification ; Bacteriological Techniques/methods ; Bacteriophages/genetics ; Bacteriophages/metabolism ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Food Microbiology/methods ; Genome, Viral/genetics ; Sequence Deletion ; Luminescent Measurements/methods ; Sensitivity and Specificity |
| Chemical Substances | Viral Proteins |
| Language | English |
| Publishing date | 2023-11-04 |
| Publishing country | Canada |
| Document type | Journal Article |
| ZDB-ID | 1111695-x |
| ISSN | 1873-7145 ; 0963-9969 |
| ISSN (online) | 1873-7145 |
| ISSN | 0963-9969 |
| DOI | 10.1016/j.foodres.2023.113665 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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