P1-08 Relative Limit of Detection Comparison for Two Rapid Detection Methods to Accurately Detect Listeria monocytogenes Analyte in Hot Dog and Deli Turkey Matrices

Sunday, July 26, 2015
Exhibit Hall (Oregon Convention Center)
Samantha Stewart , Texas Tech University , Lubbock , TX
Currey Nobles
W. Evan Chaney , Roka Bioscience, Inc. , Warren , NJ
Chanelle Adams
Erin Dreyling , Roka Bioscience, Inc. , Warren , NJ
Kendra Nightingale , Texas Tech University , Lubbock , TX
Introduction: The presence of Listeria monocytogenes in Ready-to-Eat (RTE) foods is a significant public health risk. Detection methods with enhanced sensitivity may reduce false negatives and aid in preventing contaminated product entering the food supply.

Purpose: To compare the Atlas® Listeria monocytogenes LmG2 Detection Assay to the Dupont™ BAX® System PCR Assay for detection of Listeria monocytogenes in RTE meat matrices using a relative Limit of Detection (rLOD) approach. 

Methods: Hot dog and deli turkey were bulk batch inoculated targeting 4 - 8 CFU/sample, held at 4°C for 48 - 72 h and then portioned into twenty replicate samples per matrix and method. Five un-inoculated samples were included for each method. For the Atlas®, 1:9 and 1:4 enrichment ratios were prepared in PALCAM broth + 0.02 g/l nalidixic acid and incubated at 35 ± 2°C for 24 h (25 g hot dogs) or 48 h (125 g deli turkey), respectively. For the BAX, a 1:9 dilution was prepared in UVM broth for both matrices, and incubated at 30 ± 2°C for 24 h (25 g hot dogs) or 26 h (125 g deli turkey), respectively, prior to secondary enrichment in MOPS-BLEB for 24 h prior to analysis. End-point enriched samples were 10-fold serially diluted and subjected to testing by each respective method. Culture based MPN was utilized to estimate starting concentrations.

Results: For hot dogs, the Atlas detected 20/20 positives for the 1:1000 dilution, whereas BAX detected 7/20. The rLOD value was 14.383, indicating significantly different method sensitivities (P < 0.001). Similarly, for deli turkey at the 1:10,000 dilution, Atlas detected 14/20 positives and BAX 0/20. The rLOD value was 112.441, indicating significantly different method sensitivities (P < 0.001). Un-inoculated replicates were negative for both methods. Between test Chi-square analyses at each dilution further supported rLOD analyses.

Significance: Rapid methods with enhanced sensitivity may provide more accurate test results for products with extremely low level contamination.