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Evaluation of an automated repetitive sequence-based PCR system for subtyping Enterobacter sakazakii
1Centre for Food Safety, School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland.
Insights
A new PCR method effectively subtypes Enterobacter sakazakii, a pathogen found in infant formula. This approach aids in monitoring contamination within manufacturing facilities to protect infant health.
Area of Science:
- Food safety and microbiology
- Bacterial typing and epidemiology
Background:
- Enterobacter sakazakii is a ubiquitous bacterium found in various environments.
- Infant infections are linked to contaminated powdered infant formula (PIF).
- Contamination control in PIF manufacturing is crucial for infant health and consumer confidence.
Purpose of the Study:
- To apply repetitive sequence-based PCR (rep-PCR) for subtyping Enterobacter sakazakii.
- To compare the discriminatory power of rep-PCR with pulsed-field gel electrophoresis (PFGE).
- To evaluate rep-PCR for monitoring in-process contamination in manufacturing.
Main Methods:
- A collection of well-characterized E. sakazakii isolates of diverse origins was used.
- Repetitive sequence-based PCR (rep-PCR) was employed for subtyping.
- Results were compared with existing pulsed-field gel electrophoresis (PFGE) data.
Main Results:
- Both rep-PCR and PFGE discriminated between E. sakazakii isolates.
- Rep-PCR identified 65 distinct types.
- PFGE identified 110 types with >=95% similarity, indicating rep-PCR is less discriminatory but faster.
Conclusions:
- Rep-PCR is a rapid and effective method for subtyping E. sakazakii.
- This method is valuable for monitoring contamination in food manufacturing.
- Application of rep-PCR can help reduce pathogen transmission and enhance food safety.
Abstract:
Enterobacter sakazakii is regarded as a ubiquitous organism that can be isolated from a wide range of foods and environments. Infection in at-risk infants has been epidemiologically linked to the consumption of contaminated powdered infant formula. Preventing the dissemination of this pathogen in a powdered infant formula manufacturing facility is an important step in ensuring consumer confidence in a given brand together with the protection of the health status of a vulnerable population. In this study we report the application of a repetitive sequence-based PCR typing method to subtype a previously well-characterized collection of E. sakazakii isolates of diverse origin. While both methods successfully discriminated between the collection of isolates, repetitive sequence-based PCR identified 65 types, whereas pulsed-field gel electrophoresis identified 110 types showing > or =95% similarity. The method was quick and easy to perform, and our data demonstrated the utility and value of this approach to monitor in-process contamination, which could potentially contribute to a reduction in the transmission of E. sakazakii.
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