Listeriosis is one of the most important food-borne diseases. A variety of culture and rapid methods are available for the detection of Listeria spp. in foods. Although the presence of L. innocua may indicate potential contamination with L. monocytogenes, only the latter species is pathogenic for humans. Therefore, the most adequate tests are those which specifically detect L. monocytogenes. Chromogenic media is currently the most common method used for the presumptive identification of L. monocytogenes. Some tests like those based on antigen detection are fast and easily applied, but only a few may specifically detect L. monocytogenes. Real-time polymerase chain reaction is increasingly applied in food diagnostics for the detection of L. monocytogenes due to the availability of different specific commercial test methods. Microarrays and biosensors are some examples of new technologies that might be used routinely for the detection of L. monocytogenes in foods in the future.

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A SYBR Green I based real-time PCR assay with inlA-specific oligonucleotide primers was developed for easy and rapid detection of Listeria monocytogenes in a model food that usually has a high incidence of contamination with this pathogen. Results with pure cultures and artificially contaminated chicken meat samples indicate that the PCR assay was highly specific and sensitive. The melting point analysis of the 160 bp amplified DNA fragment was different for L. monocytogenes isolates of the two major phylogenetic divisions of the species, 1 and 2. The assay was then used to survey retail ground chicken meat for contamination with L. monocytogenes. Thirty-seven samples were enriched according to the United States Department of Agriculture culture assays to detect L. monocytogenes on meat. The use and efficiency of PCR assay was examined following both primary and secondary enrichments, which were also plated on chromogenic agar for enumeration of L. monocytogenes and nonpathogenic Listeria spp. to investigate the discrepancies between culture and PCR. Overall, L. monocytogenes was detected in 75% of the samples. Primary enrichment yielded detection rates of 70% and 37% for culture and PCR, respectively. The corresponding rates for secondary enrichment were 54% and 70%, respectively. Test sensitivity is therefore influenced by the type of enrichment and is probably related not only to the limited growth of L. monocytogenes in the primary enrichment media (false-negative PCR results), but also to the high populations of nonpathogenic Listeria spp. in the secondary enrichment broths (false-negative culture results). The main challenge of rapid PCR-based detection of L. monocytogenes from food is the poor sensitivity of primary enrichment media. The improvement of enrichment conditions may help increase assay sensitivity.

The purpose of this study was to determine whether genetically similar or diverse strains of Listeria monocytogenes colonize the environment and carcasses in a single Spanish broiler abattoir over time. The study was composed of 5 surveys over a 1.5-yr period and included the monitoring of cleaning and disinfection procedures. Overall, a total of 212 samples were tested for the presence of L. monocytogenes, and 31% of the samples were found to be positive. Listeria monocytogenes was isolated from carcasses and product contact and noncontact sites in the evisceration and carcass classification areas of the abattoir. A total of 132 L. monocytogenes isolates were characterized by PCR-based serotyping and pulsed-field gel electrophoresis (PFGE) restriction analysis with the endonucleases ApaI and AscI. Molecular serotyping showed that L. monocytogenes isolates were of serotypes 1/2a and 1/2b. Isolates of serotype 1/2b (89.4%) were contaminating carcasses as well as environmental product contact and noncontact sites, whereas isolates of serotype 1/2a (10.6%) were recovered only from environmental product noncontact sites. A relatively low genetic diversity was found in this group of L. monocytogenes isolates from the abbatoir; only 14 different PFGE types (A1 to A14) were obtained. Nine pulsotypes belonging to serotype 1/2b (lineage I) were grouped in only one PFGE genetic cluster, whereas 5 pulsotypes belonging to serotype 1/2a (lineage II) were grouped into 4 PFGE genetic clusters. Two genetically related pulsotypes of serotype 1/2b (A1 and A2, 64.4% of the isolates) predominated and persisted in the abattoir. Our study indicated that a few strains of L. monocytogenes lineage I that were genetically very closely related might be specifically adapted to colonizing the evisceration zone of the abattoir and were predominant on carcasses over 1 yr. On the other hand, a genetically diverse group of lineage II strains were present in the abattoir environment, but never contaminated carcasses.