Next day Salmonella spp. detection method based on real-time PCR for meat, dairy and vegetable food products

• To compare RTi-PCR strategies for a rapid detection of Salmonella spp.
• Several parameters were evaluated to reduce the time and cost for final results.
• An 18 h-incubation coupled to a DNA extraction by boiling and RTi-PCR detected down to 2 CFU.
• It is fully compatible with the ISO standard, providing results more rapidly and cost-effectively.
• The results were confirmed in a large number of naturally contaminated food samples.

The microbiological standard for detection of Salmonella relies on several cultural steps and requires more than 5 days for final confirmation, and as consequence there is a need for an alternative rapid methodology for its detection. The aim of this study was to compare different detection strategies based on real-time PCR for a rapid and sensitive detection in an ample range of food products: raw pork and poultry meat, ready to eat lettuce salad and raw sheep milk cured cheese. Three main parameters were evaluated to reduce the time and cost for final results: the initial sample size (25 and 50 g), the incubation times (6, 10 and 18 h) and the bacterial DNA extraction (simple boiling of the culture after washing the bacterial pellet, the use of the Chelex resin, and a commercial silica column). The results obtained demonstrate that a combination of an incubation in buffered peptone water for 18 h of a 25 g-sample coupled to a DNA extraction by boiling and a real-time PCR assay detected down to 2–4 Salmonella spp. CFU per sample in less than 21 h in different types of food products. This RTi-PCR-based method is fully compatible with the ISO standard, providing results more rapidly and cost-effectively. The results were confirmed in a large number of naturally contaminated food samples with at least the same analytical performance as the reference method.