A quantitative PCR to follow-up germination and growth of Clostridium botulinum in refrigerated processed foods with extended durability
- Author
- Katrien Begyn (UGent) , Dan Li (UGent) , Frank Devlieghere (UGent) and Mieke Uyttendaele (UGent)
- Organization
- Abstract
- Introduction: Clostridium botulinum is a gram-positive, spore forming, anaerobic bacteria which produces the lethal neurotoxin. Two groups of C. botulinum are implemented in food-related intoxication (group I & group II). As group II C. botulinum are psychotropic, they play an important role in the safety of Refrigerated processed food with extended durability (REPFED). To guarantee the safety of REPFED a pasteurization (P90) is done. However, this severe heat treatment has a negative influence on the quality of the REPFEDs [1, 2]. Any alterations to this temperature should be carefully examined. At the moment there is no culture method available for selective enumeration of C. botulinum, which makes challenge test with C. botulinum difficult. To be able to follow-up germination and thus growth of C .botulinum in a food matrix a qPCR was developed. Material and methods: Two type of psychotropic C. botulinum strains were used (NCTC 8550 & NCTC 11219). DNA was extracted from cells, spores and inoculated foods using the NucleoSpin Tissue kit (Macherey-Nagel). Primers against the toxin E genes of psychotropic C. botulinum type E strains were based on Kirchner et al. (2010) and used together with Power SYBRGreen PCR MM (Invitrogen) [3]. Results: The detection limit based on pure cultures of the two C. botulinum strains (NCTC 8550 & NCTC 11219) was set at 200 CFU/mL. Based on the analyses of multiple negative controls and 8 different strains a Ct-value higher than 30 was seen as negative. Fish and fish mousse was inoculated with 500 CFU/g, which resulted in a Ct-value of 26,6 and 25,64 respectively for strains NCTC 8550 and 25,65 and 24,52 respectively for strain NCTC 11219. Discussion: The primers used here were based on the pentaplexed real-time PCR of Kirchner et al. (2010) [3]. Kirchner et al. used a Taqman based qPCR and tested the primers against 42 C. botulinum and 57 non-C. botulinum strains. The detection limit in a range of foods ranged from 103 CFU/mL to 104 CFU/mL depending on the type of food used [3]. Here, we show a lower detection limit of 500 CFU/g in fish. More research is necessary to determine the detection limit in multiple REPFEDs and to investigate the ability to follow-up germination and growth in these REPFEDs. References 1. Peck, M.W., Clostridium botulinum and the safety of refrigerated processed foods of extended durability. Trends in Food Science & Technology, 1997. 8(6): p. 186-192. 2. Peck, M.W., Clostridium botulinum and the safety of minimally heated, chilled foods: an emerging issue? Journal of Applied Microbiology, 2006. 101(3): p. 556-570. 3. Kirchner, S., et al., Pentaplexed quantitative real-time PCR assay for the simultaneous detection and quantification of botulinum neurotoxin-producing clostridia in food and clinical samples. Applied and Environmental Microbiology, 2010. 76(13): p. 4387-95.
Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-7276442
- MLA
- Begyn, Katrien, et al. “A Quantitative PCR to Follow-up Germination and Growth of Clostridium Botulinum in Refrigerated Processed Foods with Extended Durability.” Food Microbiology, 20th Conference, Abstracts, Belgian Society for Food Microbiology (BSFM), 2015.
- APA
- Begyn, K., Li, D., Devlieghere, F., & Uyttendaele, M. (2015). A quantitative PCR to follow-up germination and growth of Clostridium botulinum in refrigerated processed foods with extended durability. Food Microbiology, 20th Conference, Abstracts. Presented at the 20th Conference on Food Microbiology (BSFM 2015), Brussels, Belgium.
- Chicago author-date
- Begyn, Katrien, Dan Li, Frank Devlieghere, and Mieke Uyttendaele. 2015. “A Quantitative PCR to Follow-up Germination and Growth of Clostridium Botulinum in Refrigerated Processed Foods with Extended Durability.” In Food Microbiology, 20th Conference, Abstracts. Belgian Society for Food Microbiology (BSFM).
- Chicago author-date (all authors)
- Begyn, Katrien, Dan Li, Frank Devlieghere, and Mieke Uyttendaele. 2015. “A Quantitative PCR to Follow-up Germination and Growth of Clostridium Botulinum in Refrigerated Processed Foods with Extended Durability.” In Food Microbiology, 20th Conference, Abstracts. Belgian Society for Food Microbiology (BSFM).
- Vancouver
- 1.Begyn K, Li D, Devlieghere F, Uyttendaele M. A quantitative PCR to follow-up germination and growth of Clostridium botulinum in refrigerated processed foods with extended durability. In: Food Microbiology, 20th Conference, Abstracts. Belgian Society for Food Microbiology (BSFM); 2015.
- IEEE
- [1]K. Begyn, D. Li, F. Devlieghere, and M. Uyttendaele, “A quantitative PCR to follow-up germination and growth of Clostridium botulinum in refrigerated processed foods with extended durability,” in Food Microbiology, 20th Conference, Abstracts, Brussels, Belgium, 2015.
@inproceedings{7276442, abstract = {{Introduction: Clostridium botulinum is a gram-positive, spore forming, anaerobic bacteria which produces the lethal neurotoxin. Two groups of C. botulinum are implemented in food-related intoxication (group I & group II). As group II C. botulinum are psychotropic, they play an important role in the safety of Refrigerated processed food with extended durability (REPFED). To guarantee the safety of REPFED a pasteurization (P90) is done. However, this severe heat treatment has a negative influence on the quality of the REPFEDs [1, 2]. Any alterations to this temperature should be carefully examined. At the moment there is no culture method available for selective enumeration of C. botulinum, which makes challenge test with C. botulinum difficult. To be able to follow-up germination and thus growth of C .botulinum in a food matrix a qPCR was developed. Material and methods: Two type of psychotropic C. botulinum strains were used (NCTC 8550 & NCTC 11219). DNA was extracted from cells, spores and inoculated foods using the NucleoSpin Tissue kit (Macherey-Nagel). Primers against the toxin E genes of psychotropic C. botulinum type E strains were based on Kirchner et al. (2010) and used together with Power SYBRGreen PCR MM (Invitrogen) [3]. Results: The detection limit based on pure cultures of the two C. botulinum strains (NCTC 8550 & NCTC 11219) was set at 200 CFU/mL. Based on the analyses of multiple negative controls and 8 different strains a Ct-value higher than 30 was seen as negative. Fish and fish mousse was inoculated with 500 CFU/g, which resulted in a Ct-value of 26,6 and 25,64 respectively for strains NCTC 8550 and 25,65 and 24,52 respectively for strain NCTC 11219. Discussion: The primers used here were based on the pentaplexed real-time PCR of Kirchner et al. (2010) [3]. Kirchner et al. used a Taqman based qPCR and tested the primers against 42 C. botulinum and 57 non-C. botulinum strains. The detection limit in a range of foods ranged from 103 CFU/mL to 104 CFU/mL depending on the type of food used [3]. Here, we show a lower detection limit of 500 CFU/g in fish. More research is necessary to determine the detection limit in multiple REPFEDs and to investigate the ability to follow-up germination and growth in these REPFEDs. References 1. Peck, M.W., Clostridium botulinum and the safety of refrigerated processed foods of extended durability. Trends in Food Science & Technology, 1997. 8(6): p. 186-192. 2. Peck, M.W., Clostridium botulinum and the safety of minimally heated, chilled foods: an emerging issue? Journal of Applied Microbiology, 2006. 101(3): p. 556-570. 3. Kirchner, S., et al., Pentaplexed quantitative real-time PCR assay for the simultaneous detection and quantification of botulinum neurotoxin-producing clostridia in food and clinical samples. Applied and Environmental Microbiology, 2010. 76(13): p. 4387-95.}}, author = {{Begyn, Katrien and Li, Dan and Devlieghere, Frank and Uyttendaele, Mieke}}, booktitle = {{Food Microbiology, 20th Conference, Abstracts}}, language = {{eng}}, location = {{Brussels, Belgium}}, publisher = {{Belgian Society for Food Microbiology (BSFM)}}, title = {{A quantitative PCR to follow-up germination and growth of Clostridium botulinum in refrigerated processed foods with extended durability}}, year = {{2015}}, }