An Alternative Rapid Confirmation Method for Identifying Listeria monocytogenes from a Variety of 125 g Food Samples Within Two Days of a PCR Presumptive Positive

• Published in: Journal of food protection 2024-01, Vol.87 (1), p.100193-100193, Article 100193
• Main Authors: Carlin, Catharine R., Akins-Lewenthal, Deann, Bastin, Benjamin, Crowley, Erin, McMahon, Wendy, Ziebell, Bradley
• Format: Article
• Language: English
• Subjects: 125 g; Alternative confirmation; Fit-for-purpose; Food; Food Microbiology; L. monocytogenes; Listeria; Listeria monocytogenes - genetics; Polymerase Chain Reaction; Rapid detection; Secondary screen
• ISSN: 0362-028X; 1944-9097
• DOI: 10.1016/j.jfp.2023.100193

•L. monocytogenes was detected from a variety of foods tested at the 125 g analytical weight.•An LSB enrichment procedure was harmonized for use with three rapid method assays.•Performing a secondary rapid detection method screen reliably predicted a true positive.•L. monocytogenes colonies were identified the same day, directly from RLM agar plates.•The alternative confirmation method can provide results in 1–2 days. Cultural confirmation following detection of a Listeria monocytogenespresumptive positive can take 3–7 days to finalize; this uncertainty is a point of frustration for food producers needing to make time-sensitive disposition decisions. To address the demand for shortened time-to-results, an alternative L. monocytogenes confirmation method consisting of two components, (i) a secondary screen using a different rapid method, and (ii) concurrent cultural isolation followed by next-day colony identification was evaluated. For the study, four food matrices (hot dogs, peanut butter, frozen vegetables, and multicomponent frozen meals) were inoculated with low levels (0.36–1.39 MPN/125 g) of L. monocytogenes per the AOAC guidelines for a matrix study. Analyses were performed on 125 g test portions and started with a PCR primary screen (Bio-Rad iQ-Check Listeria monocytogenes II). Next, all enriched food samples underwent a secondary screen by bioMérieux’s GENE-UP LMO2 Real-Time PCR and VIDAS LMX ELFA along with streaking onto RAPID’L.mono Agar. Presumptive positive L. monocytogenes colonies were identified utilizing a high throughput rapid identification method (Hygiena’s BAX System L. monocytogenes Real-Time PCR assay, Neogen’s ANSR isothermal nucleic acid amplification assay, and Bruker’s MALDI Biotyper). Importantly, this study evaluated multiple commercially available options for the secondary screen (n = 2) and rapid identification (n = 3) to allow for easy adoption by testing laboratories. Overall, there was no statistically significant difference (p ≤ 0.05) between the number of L. monocytogenes-positive 125 g samples obtained by the cultural reference method and the alternative confirmation methods (regardless of which method combinations were evaluated). Additionally, this study supports that, when both the primary and secondary screen methods yield a positive result, the sample could be considered a confirmed positive for L. monocytogenes.