Listeria monocytogenes is an important food-borne pathogen and is widely tested for in food, environmental and clinical samples. Identification traditionally involved culture methods based on selective enrichment and plating followed by the characterization of Listeria spp. based on colony morphology, sugar fermentation and haemolytic properties. These methods are the gold standard; but they are lengthy and may not be suitable for testing of foods with short shelf lives. As a result more rapid tests were developed based on antibodies (ELISA) or molecular techniques (PCR or DNA hybridization). While these tests possess equal sensitivity, they are rapid and allow testing to be completed within 48 h. More recently, molecular methods were developed that target RNA rather than DNA, such as RT-PCR, real time PCR or nucleic acid based sequence amplification (NASBA). These tests not only provide a measure of cell viability but they can also be used for quantitative analysis. In addition, a variety of tests are available for sub-species characterization, which are particularly useful in epidemiological investigations. Early typing methods differentiated isolates based on phenotypic markers, such as multilocus enzyme electrophoresis, phage typing and serotyping. These phenotypic typing methods are being replaced by molecular tests, which reflect genetic relationships between isolates and are more accurate. These new methods are currently mainly used in research but their considerable potential for routine testing in the future cannot be overlooked.

Methods for the isolation and identification of Listeria spp. and Listeria monocytogenes: a review

Epidemiology of human listeriosis and seafoods.

A multi-virulence-locus sequence typing (MVLST) scheme was developed for subtyping Listeria monocytogenes, and the results obtained using this scheme were compared to those of pulsed-field gel electrophoresis (PFGE) and the published results of other typing methods, including ribotyping (RT) and multilocus sequence typing (MLST). A set of 28 strains (eight different serotypes and three known genetic lineages) of L. monocytogenes was selected from a strain collection (n > 1,000 strains) to represent the genetic diversity of this species. Internal fragments (ca. 418 to 469 bp) of three virulence genes (prfA, inlB, and inlC) and three virulence-associated genes (dal, lisR, and clpP) were sequenced and analyzed. Multiple DNA sequence alignment identified 10 (prfA), 19 (inlB), 13 (dal), 10 (lisR), 17 (inlC), and 16 (clpP) allelic types and a total of 28 unique sequence types. Comparison of MVLST with automated EcoRI-RT and PFGE with ApaI enzymatic digestion showed that MVLST was able to differentiate strains that were indistinguishable by RT (13 ribotypes; discrimination index = 0.921) or PFGE (22 profiles; discrimination index = 0.970). Comparison of MVLST with housekeeping-gene-based MLST analysis showed that MVLST provided higher discriminatory power for serotype 1/2a and 4b strains than MLST. Cluster analysis based on the intragenic sequences of the selected virulence genes indicated a strain phylogeny closely related to serotypes and genetic lineages. In conclusion, MVLST may improve the discriminatory power of MLST and provide a convenient tool for studying the local epidemiology of L. monocytogenes.

Multi-virulence-locus sequence typing of Listeria monocytogenes.

Part 1 Risks: Range of microbial risks in food processing Biofilm risks Pathogen resistance to sanitisers Aerosols as a contamination risk Consumer perceptions of risks from food. Part 2 Improving design: Improving building design Improving zoning within food processing plants Improving the design of floors Improving design of walls Improving the hygienic design of closed equipment, heating equipment, equipment handling dry materials, packaging equipment, electrical equipment, valves, pipes, pumps, sensors. Part 3 Improving hygiene management and methods: Risk assessment in hygiene management Good Manufacturing Practice (GMP) in the food industry The use of Standard Operating Procedures (SOPs) Managing risks from allergenic residues Managing contamination risks from food packaging materials Improving hygiene in food transportation Improving control of insects in food processing Improving cleaning in place (CIP) Improving cleaning out of place (COP) Improving cleaning of heat exchangers Improving cleaning of tanks Ozone decontamination in hygiene management Enzymatic cleaning in food processing Contamination routes and analysis in food processing environments Testing surface cleanability in food processing Improving the monitoring of fouling, cleaning and disinfection in closed process plant Improving surface sampling and detection of contamination Improving air sampling Testing effectiveness of disinfectants and sanitisers Traceability of disinfectants and sanitisers Improving hygiene auditing.

Handbook of Hygiene Control in the Food Industry

Genetic homogeneity among Listeria monocytogenes strains from infected patients and meat products from two geographic locations determined by phenotyping, ribotyping and PCR analysis of virulence genes

WHO-sponsored international collaborative study to evaluate methods for subtyping Listeria monocytogenes: restriction fragment length polymorphism (RFLP) analysis using ribotyping and Southern hybridization with two probes derived from L. monocytogenes chromosome

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WHO-sponsored international collaborative study to evaluate methods for subtyping Listeria monocytogenes: restriction fragment length polymorphism (RFLP) analysis using ribotyping and Southern hybridization with two probes derived from L. monocytogenes chromosome