Detection of Listeria monocytogenes in food in two days using enrichment and 5-nuclease polymerase chain reaction with end-point fluorimetry
2007
Oravcová, K.,Food Research Institute, Bratislava (Slovak Republic) | Kaclíková, E.,Food Research Institute, Bratislava (Slovak Republic) | Siekel, P.,Food Research Institute, Bratislava (Slovak Republic) | Girotti, S. | Kuchta, T.,Food Research Institute, Bratislava (Slovak Republic)
A method for the detection of Listeria monocytogenes in food in two days is presented. The method consists of a two-step enrichment, bacterial cells lysis and a closed-tube duplex 5-nuclease PCR with an internal amplification control, which is read in end-point mode in a 96-well fluorimeter. The 5-nuclease PCR has an intrinsic detection limit of 10E4 CFU per 1 ml and facilitates monitoring of false negative results caused by the failure of amplification. The complete method for the detection of L. monocytogenes in food has a detection limit of 10E0 CFU per 10 g and does not produce false positive results due to the presence of dead cells.
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