The SureFast® Listeria 3plex ONE Kit and the SureFast® PREP Bacteria Kit in a Variety of Foods and Environmental Samples: Multi-Laboratory Validation, First Action 2025.04.

内容摘要

The SureFast® Listeria 3plex ONE Kit uses the real-time polymerase chain reaction (PCR) technique for multiplex detection of Listeria species and Listeria monocytogenes in foods and environmental samples. The SureFast Listeria 3plex ONE Kit can also be used with the SureFast PREP Bacteria Kit to provide a second DNA extraction option and can be analyzed on multiple thermocyclers. The kit was evaluated using 25 g test portions in an unpaired study design for a single matrix, ready-to-eat smoked deli turkey, as part of a harmonized validation protocol with MicroVal Certification. The matrix was compared to the International Organization for Standardization Technical (ISO) 11290-1:21017 reference standard; 17 participants from 16 laboratories located within Austria, Germany, Italy, Spain, and the United States were solicited for the multi-laboratory validation (MLV), with only 10 participants submitting data. Three levels of contamination were evaluated: an uninoculated control level (0 colony-forming units (CFU)/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-10 CFU/test portion). Statistical analysis was conducted according to the Probability of Detection (POD) statistical model. Results obtained for the low inoculum level test portions produced a dLPOD value with 95% confidence interval of -0.01 (-0.04, 0.02). The dLPOD results indicate equivalence between the candidate method and reference method for the matrix evaluated and the method demonstrated acceptable multi-laboratory reproducibility as determined in the MLV. False positive and false negative rates were determined for the matrix and produced values of 0.0% and 0.8%, respectively. Based on the data generated, the method demonstrated equivalence to the ISO reference method in a MLV study. The SureFast ONE 3plex method is a rapid and reliable alternative for the multiplex detection of Listeria species and Listeria monocytogenes in a broad range of foods and environmental surfaces.