Comparative evaluation of impedanciometry combined with chromogenic agars or RNA hybridization and real-time PCR methods for the detection of L. monocytogenes in dry-cured ham

Abstract

Listeria monocytogenes is an important foodborne pathogen of particular relevance in “Ready To Eat” products. Food producers require rapid methods to detect L. monocytogenes, since the reference method (ISO 11290-1) is laborious, lengthy and costly. The aim of this study was to evaluate three alternative methods to detect L. monocytogenes in dry-cured ham following the ISO 16140-2:2016 standard: (A) impedance measurement followed by plating onto chromogenic agars; (B) impedance measurement followed by RNA hybridization, and (C) real-time PCR.Inclusivity and exclusivity were evaluated. The limits of detection 50 (LOD50) and the relative limits of detection (RLOD) were obtained by analysing dry-cured ham samples inoculated with L. monocytogenes at three different levels of contamination. The sensitivity study of alternative methods, as well as the relative specificity (SP), sensitivity (SE), and Kappa Cohen's index were calculated analysing 93 samples of sliced dry-cured ham. The inclusivity and exclusivity tests of three methods showed no interference in pathogen detection. LOD50 were very low for the three methods evaluated (<1 cfu/25 g dry-cured ham). The RLOD values of the three alternative methods were below the acceptability limit established by ISO 16140. For methods A and C, good results were obtained in the sensitivity study, as well as in the SP and SE. However, method B showed poorer results in the sensitivity study, along with lower results for SP (99.7%) and SE (79.6%), due to the occurrence of false positives and negatives in samples with presence of other Listeria spp.Methods A and C were considered to be a thoroughly appropriate control tool for use in the meat industry to improve the detection of L. monocytogenes.