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Published on: February 1, 2014
Detection of enterotoxigenic Clostridium perfringens with a duplex PCR
E Augustynowicz1, A Gzyl1, J Ślusarczyk1
1Department of Sera and Vaccines Evaluation, National Institute of Hygiene, 24 Chocimska Street, 00-791 Warsaw, Poland.
Journal of Medical Microbiology
|February 28, 2002
Summary
Detecting enterotoxigenic Clostridium perfringens using duplex PCR is effective. This method aids in identifying food poisoning-related strains, overcoming limitations of traditional serological tests due to poor sporulation.
Area of Science:
- Microbiology
- Food Safety
- Molecular Diagnostics
Background:
- Clostridium perfringens is a significant cause of foodborne illness.
- Accurate identification of enterotoxigenic strains is crucial for public health.
- Traditional methods for detecting enterotoxin production can be unreliable due to poor sporulation in fresh isolates.
Purpose of the Study:
- To evaluate a duplex PCR method for detecting Clostridium perfringens phospholipase C (plc) and enterotoxin (cpe) genes.
- To compare PCR results with in-vitro enterotoxin synthesis confirmed by serological testing.
- To assess the utility of PCR for diagnosing enterotoxigenic C. perfringens in food poisoning-related isolates.
Main Methods:
- Developed and applied two sets of primers for a single-reaction duplex PCR targeting plc and cpe genes.
- Tested 64 C. perfringens isolates, predominantly linked to food poisoning outbreaks.
- Performed in-vitro enterotoxin synthesis testing using serological methods after inducing sporulation.
Main Results:
- Out of 64 isolates, 26 were confirmed as enterotoxigenic.
- An additional 16 isolates were classified as potentially enterotoxigenic, as serological tests did not confirm enterotoxin production.
- Duplex PCR identified enterotoxigenic strains more reliably than serological analysis alone.
Conclusions:
- Duplex PCR is a valuable tool for the rapid and accurate diagnosis of enterotoxigenic Clostridium perfringens.
- This molecular method overcomes the limitations of serological testing, which can yield false negatives due to poor sporulation in fresh bacterial cultures.
- The findings support the use of PCR in routine diagnostics for foodborne pathogens.
