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A novel real‐time PCR‐based method for the detection of Listeria monocytogenes in food
Author(s) -
Oravcová K.,
Kuchta T.,
Kaclíková E.
Publication year - 2007
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2007.02234.x
Subject(s) - listeria monocytogenes , microbiology and biotechnology , biology , listeria , bacteria , polymerase chain reaction , real time polymerase chain reaction , food microbiology , virology , gene , genetics
Aims: A new real‐time PCR‐based method was developed for the detection of Listeria monocytogenes in food. Methods and Results: A two‐step enrichment involving a 24‐h incubation in half‐Fraser broth followed by a 6‐h subculture in Fraser broth was used, followed by cell lysis and real‐time PCR with primers and a TaqMan probe previously developed in our laboratory. When the method was evaluated with 144 naturally contaminated food samples, 44 were detected as positive by the PCR‐based method and 42 by the standard method EN ISO 11290‐1. With 61 food samples artificially contaminated at a level of 10 0 CFU per 25 g, 61 and 58 positive samples were detected by the respective methods. Conclusions: The developed real‐time PCR‐based method facilitated the detection of L. monocytogenes in food on the next day after the sample reception, with a reduction of false‐positive results because of dead bacterial cells and false‐negative results because of PCR inhibitors. Significance and Impact of the Study: The method can be used for L. monocytogenes detection in food as a faster alternative to current methods.