Scientific journal
Journal of Food and Nutrition Research
Summary No. 1 / 2007
ORAVCOVÁ, K. - KACLÍKOVÁ, E. - SIEKEL, P. - GIROTTI, S. - KUCHTA, T.
Detection of Listeria monocytogenes in food in two days using enrichment and 5’-nuclease polymerase chain reaction with end-point fluorimetry
Journal of Food and Nutrition Research, 46, 2007, No. 1, s. 35-38
Tomáš Kuchta, Department of Microbiology and Molecular Biology, VÚP Food Research Institute, Priemyselná 4, P. O. Box 25, SK-824 75 Bratislava 26, Bratislava, Slovakia. E-mail: kuchta@vup.sk
Summary: 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 104 CFU.ml-1 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 100 CFU per 10 g and does not produce false positive results due to the presence of dead cells.
Keywords: Listeria monocytogenes; polymerase chain reaction; rapid method; end-point fluorimetry
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