Rapid Purification of Salmonella DNA in Minced Meat and Detection by Real-time PCR

G. Jenikova, Annette Nygaard Jensen, K. Demnerova, Jeffrey Hoorfar

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Four rapid and simple DNA purification and sample treatment protocols were evaluated for detection of Salmonella enterica in spiked minced meat, using a fluorogenic 5' nuclease (TaqMan) PCR assay in an ABI-Prism 7700 Sequence Detector. The detection limit with the single separation treatment of DNeasy was found to be 6-8 CFU in just 19 end-point fluorescence (C-t) values, while this was 22 C-t for a combination of DNeasy and BactXtractor. Extraction by DNeasy resulted in C-t <22, indicating a detection limit of <10 Salmonella cells per 25 g, when the samples were inoculated with Salmonella before the overnight preenrichment. The method is currently being adapted to a BioRobot 3000 platform. However, the use of paramagnetic beads (DNA Direct) resulted in poor and variable detection limit.
Original languageEnglish
JournalJournal of Rapid Methods and Automation in Microbiology
Volume9
Issue number3
Pages (from-to)135-141
ISSN1060-3999
Publication statusPublished - 2001

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