Purpose: An independent study was conducted at ADRIA, to validate this new method in comparison to the ISO 11290-1 standard, as part of the NF Validation approval process and according to the ISO 16140 standard
Methods: The alternative method includes a single step enrichment in 24 LEB supplemented with both 24 LEB Buffer and selective supplements for 22-26h at 37°C. After DNA extraction, PCR is run performed using the PikoReal instrument.
Results: 339 food and environmental samples were analyzed for relative accuracy, sensitivity and specificity. The results demonstrate equivalent performance between the alternative and ISO 11290-1 methods. Depending on the tested (matrix/strain) pairs, the relative detection limits of the alternative method vary from 0.2 to 1.2 CFU/25 g, those of the ISO standard vary from 0.2 to 1.1 CFU/25 g. The selectivity and the specificity of the alternative method were assessed by testing 50 target strains and 30 non target strains. The alternative method was also evaluated in a ring trial involving 10 laboratories. The results of the calculated accuracy, accordance, concordance and odds ratio clearly show that the alternative method precision is equivalent to the ISO 11290-1 reference method.
Significance: The SureTect™ Listeria monocytogenes Real-Time PCR Assay is a reliable method for Listeria monocytogenes detection in food and environmental samples, and offers important economic savings by reducing time to result and handling time.