Restriction Fragment Length Polymorphisms Detected with Novel DNA Probes Differentiate among Diverse Lineages of Serogroup 4
            <i>Listeria monocytogenes</i>
            and Identify Four Distinct Lineages in Serotype 4b

Tran, Huyen L.; Kathariou, Sophia

doi:10.1128/aem.68.1.59-64.2002

Cited by 19 publications

(10 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To date, only one RFLP that differentiates ECII from other serotype 4b strains has been identified. This polymorphism was detected with a DNA probe derived from the putative mannitol permease locus of L. monocytogenes, which differentiated among several lineages within serotype 4b (27).…”

Section: Discussionmentioning

confidence: 99%

Genetic Markers Unique to Listeria monocytogenes Serotype 4b Differentiate Epidemic Clone II (Hot Dog Outbreak Strains) from Other Lineages

Evans

Swaminathan

Graves

et al. 2004

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

A small number of closely related strains of Listeria monocytogenes serotype 4b, designated epidemic clone I (ECI), have been implicated in numerous outbreaks of food-borne listeriosis described during the past two decades in Europe and North America. In 1998 to 1999, a multistate outbreak traced to contaminated hot dogs involved a different strain type of serotype 4b, with genetic fingerprints rarely encountered before. In spite of the profound economic and public health impact of this outbreak, the implicated bacteria (designated epidemic clone II [ECII]) have remained poorly characterized genetically, and nucleotide sequences specific for these strains have not been reported. Using genome sequence information, PCR, and Southern blots, we identified DNA fragments which appeared to be either absent or markedly divergent in the hot dog outbreak strains but conserved among other serotype 4b strains. PCR with primers derived from these fragments as well as Southern blots with the amplicons as probes readily differentiated ECII from other serotype 4b strains. The serotype 4b-specific region harboring these fragments was adjacent to inlA, which encodes a well-characterized virulence determinant. The findings suggest that ECII strains have undergone divergence in portions of a serotype-specific region that is conserved in other serotype 4b strains. Although the mechanisms that drive this divergence remain to be identified, DNA-based tools from this region can facilitate the detection and further characterization of strains belonging to this lineage.

show abstract

Section: Discussionmentioning

confidence: 99%

Genetic Markers Unique to Listeria monocytogenes Serotype 4b Differentiate Epidemic Clone II (Hot Dog Outbreak Strains) from Other Lineages

Evans

Swaminathan

Graves

et al. 2004

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…detection in food and environmental samples have been reported (1,4). Molecular methods used for the genetic identification and characterization of Listeria isolates include PCR with primers specific for characteristic genes (6,23,31), restriction fragment length polymorphism analysis (42,44), amplified fragment length polymorphism analysis (34), random amplification of polymorphic DNA (47), and pulsed-field gel electrophoresis (25).…”

Section: The Genus Listeria Consists Of Six Species: L Monocytogenesmentioning

confidence: 99%

Identification ofListeriaSpecies by Microarray-Based Assay

et al. 2002

View full text Add to dashboard Cite

We have developed a rapid microarray-based assay for the reliable detection and discrimination of six species of the Listeria genus: L. monocytogenes, L. ivanovii, L. innocua, L. welshimeri, L. seeligeri, and L. grayi. The approach used in this study involves one-tube multiplex PCR amplification of six target bacterial virulence factor genes (iap, hly, inlB, plcA, plcB, and clpE), synthesis of fluorescently labeled single-stranded DNA, and hybridization to the multiple individual oligonucleotide probes specific for each Listeria species and immobilized on a glass surface. Results of the microarray analysis of 53 reference and clinical isolates of Listeria spp. demonstrated that this method allowed unambiguous identification of all six Listeria species based on sequence differences in the iap gene. Another virulence factor gene, hly, was used for detection and genotyping all L. monocytogenes, all L. ivanovii, and 8 of 11 L. seeligeri isolates. Other members of the genus Listeria and three L. seeligeri isolates did not contain the hly gene. There was complete agreement between the results of genotyping based on the hly and iap gene sequences. All L. monocytogenes isolates were found to be positive for the inlB, plcA, plcB, and clpE virulence genes specific only to this species. Our data on Listeria species analysis demonstrated that this microarray technique is a simple, rapid, and robust genotyping method that is also a potentially valuable tool for identification and characterization of bacterial pathogens in general.

show abstract

“…Of these, serotype 4b may be the most virulent; it accounts for nearly all of the outbreaks of human food-borne and perinatal listeriosis even though it is not found as commonly in foods or the environment as other serotypes (18,32,43,57). Moreover, serotype 4b strains isolated from outbreaks comprise two genetically uniform subtypes compared to the diversity of serotype 4b strains isolated from the environment, suggesting that subpopulations within the 4b serotype may comprise one or more epidemic clones (54,61).…”

mentioning

confidence: 99%

“…Previous population genetic analyses and comparative genotyping studies have suggested that significant genome diversity exists among the different serotypes (1,3,7,26,27,44,47,54,57,58,61). Comparative genome analyses using subtractive libraries suggest that as much as 5% of the serotype 4b genome is not present in serotype 1/2a strains and that the patterns of diversity are congruent with serotype distribution (30).…”

mentioning

confidence: 99%

Genome Diversification in Phylogenetic Lineages I and II ofListeria monocytogenes: Identification of Segments Unique to Lineage II Populations

Zhang¹,

Ju²,

Nietfeldt³

et al. 2003

J Bacteriol

View full text Add to dashboard Cite

Thirteen different serotypes of Listeria monocytogenes can be distinguished on the basis of variation in somatic and flagellar antigens. Although the known virulence genes are present in all serotypes, greater than 90% of human cases of listeriosis are caused by serotypes 1/2a, 1/2b, and 4b and nearly all outbreaks of food-borne listeriosis have been caused by serotype 4b strains. Phylogenetic analysis of these three common clinical serotypes places them into two different lineages, with serotypes 1/2b and 4b belonging to lineage I and 1/2a belonging to lineage II. To begin examining evolution of the genome in these serotypes, DNA microarray analysis was used to identify lineage-specific and serotype-specific differences in genome content. A set of 44 strains representing serotypes 1/2a, 1/2b, and 4b was probed with a shotgun DNA microarray constructed from the serotype 1/2a strain 10403s. Clones spanning 47 different genes in 16 different contiguous segments relative to the lineage II 1/2a genome were found to be absent in all lineage I strains tested (serotype 4b and 1/2b) and an additional nine were altered exclusively in 4b strains. Southern hybridization confirmed that conserved alterations were, in all but two loci, due to absence of the segments from the genome. Genes within these contiguous segments comprise five functional categories, including genes involved in synthesis of cell surface molecules and regulation of virulence gene expression. Phylogenetic reconstruction and examination of compositional bias in the regions of difference are consistent with a model in which the ancestor of the two lineages had the 1/2 somatic serotype and the regions absent in the lineage I genome arose by loss of ancestral sequences.Listeria monocytogenes is a ubiquitous gram-positive bacterium that can cause life-threatening infections including meningitis, septicemia, abortion, and fetal death. Its primary route of transmission to humans is through contaminated food and despite the relatively low incidence of listeriosis in humans, outbreaks of listeriosis often have high associated morbidity, particularly among pregnant women, unborn fetuses, and immunocompromised individuals (24). These characteristics, coupled with its physiological durability and its ubiquitous distribution in nature have propelled L. monocytogenes to the forefront of food safety research and the regulatory arena.Pathogenesis of listeriosis is a consequence of the organism's ability to invade and replicate within several different cell types in mammalian tissues, including intestinal, liver, and neural tissues. During the course of food-borne infections, the bacteria penetrate the intestinal lining through cell invasion and translocation and use the lymphatic system as a conduit to reach the main target tissues within the liver and spleen (41). Prolonged replication in the liver, facilitated by depressed cell mediated immunity, is thought to be an antecedent to spread of the bacteria to brain tissue and breach of the placental barrier in pregnant wo...

show abstract

Restriction Fragment Length Polymorphisms Detected with Novel DNA Probes Differentiate among Diverse Lineages of Serogroup 4 Listeria monocytogenes and Identify Four Distinct Lineages in Serotype 4b

Cited by 19 publications

References 36 publications

Genetic Markers Unique to Listeria monocytogenes Serotype 4b Differentiate Epidemic Clone II (Hot Dog Outbreak Strains) from Other Lineages

Genetic Markers Unique to Listeria monocytogenes Serotype 4b Differentiate Epidemic Clone II (Hot Dog Outbreak Strains) from Other Lineages

Identification ofListeriaSpecies by Microarray-Based Assay

Genome Diversification in Phylogenetic Lineages I and II ofListeria monocytogenes: Identification of Segments Unique to Lineage II Populations

Contact Info

Product

Resources

About