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Enrichment and Detection of Clostridium perfringens Toxinotypes in Retail Food Samples
Published on: October 18, 2019
Rapid detection of enterotoxigenic Clostridium perfringens in meat samples using immunomagnetic separation polymerase
Zheng-You Yang1, Won-Bo Shim, Kyeong-Yeol Kim
1Department of Microbiology, College of Life Science, Key Laboratory for Agriculture Microbiology, Shandong Agricultural University, Taian, China.
Abstract:
Rapid detection of enterotoxigenic Clostridium perfringens in meat samples was accomplished with an immunomagnetic separation polymerase chain reaction (IMS-PCR). First, a monoclonal antibody (mAb) specific to C. perfringens was generated. The antibody showed strong binding to C. perfringens and no binding to non- Clostridia bacteria, except a weak cross-reaction to Staphylococcus aureus based on the enzyme-linked immunosorbent assay (ELISA). Then, magnetic beads were coated with the mAb, and the IMS-PCR system was developed. With the optimized conditions, the IMS-PCR assay was capable of detecting as few as 10 colony forming units (CFU)/g of C. perfringens cells in the meat sample within 10 h. Of the 116 collected samples (26 chicken samples, 20 beef samples, 30 pork samples, 20 fish samples, and 20 processed meat samples) examined with IMS-PCR, 36 (31%) were C. perfringens -positive samples and 2 (1.7%) were enterotoxigenic C. perfringens -positive samples. The IMS-PCR results gave a good agreement with the results obtained by conventional culture methods. In comparison to conventional culture methods, the IMS-PCR is a rapid and specific method and has potential use as a screening tool for enterotoxigenic C. perfringens in food samples.
Insights
A new immunomagnetic separation polymerase chain reaction (IMS-PCR) method rapidly detects enterotoxigenic Clostridium perfringens in meat. This rapid detection tool offers significant potential for food safety screening.
Area of Science:
- Food microbiology
- Molecular biology
- Immunology
Background:
- Clostridium perfringens is a significant foodborne pathogen.
- Accurate and rapid detection methods are crucial for food safety.
Purpose of the Study:
- To develop and validate a rapid immunomagnetic separation polymerase chain reaction (IMS-PCR) assay for detecting enterotoxigenic Clostridium perfringens in meat samples.
Main Methods:
- Generation of a specific monoclonal antibody (mAb) against C. perfringens.
- Coating magnetic beads with the mAb to create an IMS system.
- Integration of IMS with PCR for rapid detection.
Main Results:
- The IMS-PCR assay detected as few as 10 CFU/g of C. perfringens within 10 hours.
- Out of 116 meat samples, 36 tested positive for C. perfringens and 2 for enterotoxigenic strains.
- Results showed good agreement with conventional culture methods.
Conclusions:
- The developed IMS-PCR method is a rapid, specific, and sensitive tool for detecting enterotoxigenic C. perfringens in food.
- This method has potential as a screening tool to enhance food safety protocols.
