Evaluation of Larger Test Portion Sizes for Escherichia coli Shiga Toxin Producer (STEC) on the Detection by Immunomagnetic Separation and Real-Time PCR in Meat and Vegetables - VetAgro Sup
Article Dans Une Revue Food Analytical Methods Année : 2023

Evaluation of Larger Test Portion Sizes for Escherichia coli Shiga Toxin Producer (STEC) on the Detection by Immunomagnetic Separation and Real-Time PCR in Meat and Vegetables

Résumé

The aims of this study were (i) to evaluate the performance of the Assurance ® GDS method combining immunomagnetic separation and real-time PCR for STEC detection in pooled samples (up to 375g) of vegetables and meat, and (ii) to compare its performances to that of the reference method ISO/TS-13136:2012 (25 g sample size) in artificially contaminated samples. The alternative Assurance ® GDS method includes enrichment in proprietary broth at 41.5°C for 10 h followed by primary screening of TOP 7 STEC containing eae , stx genes and O157:H7 markers using MPX-Top 7 (IMS+PCR), secondary screening for serogroup identification using MPX-ID/EHEC-ID (IMS+PCR). And finally, cultural confirmation from same IMS beads on two selective agars is set up. For reference method, the enrichment was performed in BPW at 41.5°C for 18 h. A total of 120 samples of meat and vegetables, including 20 uninoculated and 100 samples spiked with stressed cells (<12 CFU/25 or 375g) of STEC, were analyzed using both methods. Our results showed that the Assurance ® GDS method LOD 50 ranged from 0.12 CFU/375 to 0.522 CFU/375 g and the LOD 95 from 0.518 CFU/375 to 2.257 CFU/375 g. These data are similar to the LOD50 data of the reference method.
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hal-04641901 , version 1 (23-07-2024)

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Marion Bouvier-Crozier, M. Canizares, B. Hamadou, M. Guenser, Delphine Thevenot-Sergentet. Evaluation of Larger Test Portion Sizes for Escherichia coli Shiga Toxin Producer (STEC) on the Detection by Immunomagnetic Separation and Real-Time PCR in Meat and Vegetables. Food Analytical Methods, 2023, 16 (7), pp.1271-1282. ⟨10.1007/s12161-023-02505-5⟩. ⟨hal-04641901⟩
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