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Detection of salmonella in meat: Comparative and interlaboratory validation of a noncomplex and cost-effective pre-PCR protocol
DTU Technical University of Denmark, Denmark.ORCID iD: 0000-0001-6492-1245
2012 (English)In: Journal of AOAC International, ISSN 1060-3271, E-ISSN 1944-7922, Vol. 95, no 1, p. 100-104Article in journal (Refereed) Published
Abstract [en]

Cost-effective and rapid monitoring of Salmonella in the meat production chain can contribute to food safety. The objective of this study was to validate an easy-to-use pre-PCR sample preparation method based on a simple boiling protocol for screening of Salmonella in meat and carcass swab samples using a real-time PCR method. The protocol included incubation in buffered peptone water, centrifugation of an aliquot, and a boiling procedure. The validation study included comparative and interlaboratory trials recommended by the Nordic Organization for Validation of Alternative Microbiological Methods (NordVal). The comparative trial was performed against a reference method (NMKL 187, 2007) and a PCR method previously approved by NordVal with a semiautomated magnetic bead-based DNA extraction step using 122 artificially contaminated samples. The LOD was found to be 3.0, 3.2, and 3.4 CFU/sample for the boiling, magnetic bead-based, and NMKL 187 methods, respectively. When comparing the boiling method with the magnetic beads, the relative accuracy (AC), relative sensitivity (SE), and relative specificity (SP) were 98, 102, and 98%, respectively (Cohen's kappa index 0.95). When comparing results obtained by the boiling to the culture-based method, the AC, SE, and SP were found to be 98, 102, and 98%, respectively (kappa index 0.93). In the interlaboratory trial including valid results from 11 laboratories, apart from two falsepositive samples by the boiling method combined with PCR, no deviating results were obtained (SP, SE, and AC were 100, 95, and 97%, respectively). This test is under implementation by the Danish meat industry, and can be useful for screening of large number of samples in the meat production, especially for fast release of minced meat with a short shelf life. © 2012 Publishing Technology.

Place, publisher, year, edition, pages
2012. Vol. 95, no 1, p. 100-104
Keywords [en]
Culture-based methods, DNA extraction, Kappa index, Magnetic beads, Meat industry, Meat production, Microbiological methods, Minced meat, Number of samples, PCR method, Rapid monitoring, Real-time PCR method, Reference method, Relative accuracy, Relative sensitivity, Sample preparation methods, Semi-automated, Shelf life, Validation study, Centrifugation, Cost effectiveness, DNA sequences, Meats, Phase transitions, Polymerase chain reaction, Salmonella, Screening, Thermal processing (foods), bacterial DNA, coloring agent, dyes, reagents, indicators, markers and buffers, peptone, animal, article, cattle, chemistry, clinical trial, comparative study, cost benefit analysis, culture medium, economics, isolation and purification, laboratory diagnosis, limit of detection, meat, methodology, microbiology, multicenter study, real time polymerase chain reaction, reproducibility, Salmonella typhimurium, standard, swine, Animals, Coloring Agents, Cost-Benefit Analysis, Culture Media, DNA, Bacterial, False Positive Reactions, Indicators and Reagents, Peptones, Real-Time Polymerase Chain Reaction, Reference Standards, Reproducibility of Results
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:ri:diva-39022DOI: 10.5740/jaoacint.11-093Scopus ID: 2-s2.0-84860280331OAI: oai:DiVA.org:ri-39022DiVA, id: diva2:1324773
Available from: 2019-06-14 Created: 2019-06-14 Last updated: 2019-06-14Bibliographically approved

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Löfström, Charlotta

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