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 language | English |
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Pages (from-to) | 23-29 |
Number of pages | 7 |
Journal | Food Analytical Methods |
Volume | 2 |
Issue number | 1 |
DOIs | |
Publication status | Published - Mar 2009 |
Externally published | Yes |
Keywords
- Filtration
- Hot-smoked salmon
- Immunomagnetic separation
- Listeria monocytogenes
- Real-time PCR