Detection of Salmonella enterica in food using two-step enrichment and real-time polymerase chain reaction.
OBJECTIVE
A new real-time polymerase chain reaction-based method was developed for the detection of Salmonella enterica in food.


RESULTS
The method consisted of a novel two-step enrichment involving overnight incubation in buffered peptone water and a 5-h subculture in Rappaport-Vassiliadis medium, lysis of bacterial cells and a Salmonella-specific 5'-nuclease real-time PCR with an exogenous internal amplification control. Because a two-step enrichment was used, the detection limit for dead S. enterica cells in artificially contaminated ice cream and salami samples was high at 10(7 )CFU (25 g)(-1), eliminating potential false-positive results. When the method was evaluated with a range of 100 naturally contaminated food samples, three positive samples were detected by both the real-time PCR-based method and by the standard microbiological method, according to EN ISO 6579. When the real-time PCR-based method was evaluated alongside the standard microbiological method according to EN ISO 6579 with 36 food samples artificially contaminated at a level of 10(0 )CFU (25 g)(-1), identical results were obtained from both methods.


CONCLUSIONS
The real-time PCR-based method involving a two-step enrichment produced equivalent results to EN ISO 6579 on the day after sample receipt.


CONCLUSIONS
The developed method is suitable for rapid detection of S. enterica in food.
Krascsenicsová
K
K
Department of Microbiology and Molecular Biology, Food Research Institute, Bratislava, Slovakia.
Piknová
L
L
Kaclíková
E
E
Kuchta
T
T
D005516:Food Microbiology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D019779:Salmonella enterica / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors