Detection of Salmonella enterica in Meat in Less than 5 Hours by a Low-Cost and Noncomplex Sample Preparation Method

Abstract

Salmonella is recognized as one of the most important foodborne bacteria and has wide health and socioeconomic impacts worldwide. Fresh pork meat is one of the main sources of Salmonella, and efficient and fast methods for detection are therefore necessary. Current methods for Salmonella detection in fresh meat usually include Ͼ16 h of culture enrichment, in a few cases Ͻ12 h, thus requiring at least two working shifts. Here, we report a rapid (Ͻ5 h) and high-throughput method for screening of Salmonella in samples from fresh pork meat, consisting of a 3-h enrichment in standard buffered peptone water and a real-time PCR-compatible sample preparation method based on filtration, centrifugation, and enzymatic digestion, followed by fast-cycling real-time PCR detection. The method was validated in an unpaired comparative study against the Nordic Committee on Food Analysis (NMKL) reference culture method 187. Pork meat samples (n ϭ 140) were either artificially contaminated with Salmonella at 0, 1 to 10, or 10 to 100 CFU/25 g of meat or naturally contaminated. Cohen's kappa for the degree of agreement between the rapid method and the reference was 0.64, and the relative accuracy, sensitivity, and specificity for the rapid method were 81.4, 95.1, and 97.9%, respectively. The 50% limit of detections (LOD 50 s) were 8.8 CFU/25 g for the rapid method and 7.7 CFU/25 g for the reference method. Implementation of this method will enable faster release of Salmonella low-risk meat, providing savings for meat producers, and it will help contribute to improved food safety.IMPORTANCE While the cost of analysis and hands-on time of the presented rapid method were comparable to those of reference culture methods, the fast product release by this method can provide the meat industry with a competitive advantage. Not only will the abattoirs save costs for work hours and cold storage, but consumers and retailers will also benefit from fresher meat with a longer shelf life. Furthermore, the presented sample preparation might be adjusted for application in the detection of other pathogenic bacteria in different sample types.KEYWORDS Salmonella, food safety, foodborne pathogens, fresh meat, rapid tests, real-time PCR, sample preparation, validation S almonella is a global foodborne pathogen responsible for approximately 30% of foodborne outbreaks in the United States and 23% in the European Union (1, 2). Pork meat remains an important source of human salmonellosis, which is why the absence of Salmonella is confirmed at critical control points at abattoirs (2). However, the reference culture-based methods require several days to obtain a test result (3, 4), thereby postponing the market release of fresh meat.

Combination of Immunomagnetic Separation and Real‐Time Recombinase Polymerase Amplification (IMS‐qRPA) for Specific Detection of Listeria monocytogenes in Smoked Salmon Samples

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“…FP: positive by the alternative method not confirmed as such while the reference method obtained a negative result). The reclassification of the deviations was performed as described previously (Fachmann et al., ; NordVaL, ).…”

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Antimicrobial peptides from the edible insect Musca domestica and their preservation effect on chilled pork

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