Journal article
Rapid Purification of Salmonella DNA in Minced Meat and Detection by Real-time PCR
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.
Language: | English |
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Year: | 2001 |
Pages: | 135-141 |
ISSN: | 17454581 and 10603999 |
Types: | Journal article |
DOI: | 10.1111/j.1745-4581.2001.tb00238.x |
ORCIDs: | Jensen, Annette Nygaard and Hoorfar, Jeffrey |