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 language | English |
|---|---|
| Journal | Journal of Rapid Methods and Automation in Microbiology |
| Volume | 9 |
| Issue number | 3 |
| Pages (from-to) | 135-141 |
| ISSN | 1060-3999 |
| Publication status | Published - 2001 |