Abstract
This work evaluated the application of filtration and immunomagnetic separation (IMS) as sample pretreatments for use in combination with real-time polymerase chain reaction (PCR) to detect and quantify Listeria monocytogenes in hot-smoked salmon. Salmon was artificially inoculated with L. monocytogenes at levels ranging from 8 × 100 to 8 × 105 cfu/g of sample, and homogenates obtained from these samples were filtered to recover bacterial cells without a pre-enrichment step. High recovery of bacterial cells was achieved using standard coffee filters. IMS significantly reduced the co-extraction of PCR inhibitors present in the samples to increase the assay sensitivity with regression line parameters applicable for quantification. The limit of detection and quantification were equal to 2 × 101–4 × 101 and 2 × 102 cfu/g of sample, respectively. The entire detection procedure could be completed within 3.5 h. This study demonstrated that coupling filtration and IMS with real-time PCR has contributed to improve the sensitivity of L. monocytogenes detection from hot-smoked salmon.
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Acknowledgments
This study was financially supported partially by the Research Council of Norway, grant no. 173973/I10. We gratefully thank Liv Marit Rørvik and Cudjoe Kofitsyo for their skilful contribution to this work.
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Duodu, S., Mehmeti, I., Holst-Jensen, A. et al. Improved Sample Preparation for Real-Time PCR Detection of Listeria monocytogenes in Hot-Smoked Salmon using Filtering and Immunomagnetic Separation Techniques. Food Anal. Methods 2, 23–29 (2009). https://doi.org/10.1007/s12161-008-9043-2
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DOI: https://doi.org/10.1007/s12161-008-9043-2