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Improved Sample Preparation for Real-Time PCR Detection of Listeria monocytogenes in Hot-Smoked Salmon using Filtering and Immunomagnetic Separation Techniques

Food Analytical Methods volume 2, pages 23–29 (2009)Cite this article

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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Authors and Affiliations

  1. Section for Food Bacteriology and GMO, National Veterinary Institute, Ullevaalsveien 68, P.O. Box 750, Sentrum, 0106, Oslo, Norway

    Samuel Duodu, Ibrahim Mehmeti, Arne Holst-Jensen & Semir Loncarevic

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  1. Samuel Duodu
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  2. Ibrahim Mehmeti
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  3. Arne Holst-Jensen
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  4. Semir Loncarevic
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Correspondence to Samuel Duodu.

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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

Keywords

  • Listeria Monocytogenes
  • Filtration
  • Immunomagnetic Separation
  • Real-Time PCR
  • Hot-Smoked Salmon