Improved sample preparation for real-time PCR detection of listeria monocytogenes in hot-smoked salmon using filtering and immunomagnetic separation techniques

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

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

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∈×∈10 0 to 8∈×∈ 10 5 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∈×∈10 1-4∈×∈10 1 and 2∈×∈10 2 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.

Original languageEnglish
Pages (from-to)23-29
Number of pages7
JournalFood Analytical Methods
Volume2
Issue number1
DOIs
Publication statusPublished - Mar 2009
Externally publishedYes

Keywords

  • Filtration
  • Hot-smoked salmon
  • Immunomagnetic separation
  • Listeria monocytogenes
  • Real-time PCR

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