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A novel real-time PCR-based method for the detection of Listeria monocytogenes in food

2007; Oxford University Press; Volume: 45; Issue: 5 Linguagem: Inglês

10.1111/j.1472-765x.2007.02234.x

1472-765X

Katarí­na Oravcová, T. Kuchta, Eva Kaclı́ková,

Biosensors and Analytical Detection

A new real-time PCR-based method was developed for the detection of Listeria monocytogenes in food.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.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.The method can be used for L. monocytogenes detection in food as a faster alternative to current methods.