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Subtyping of Listeria monocytogenes isolates by actA gene sequencing, PCR-fingerprinting, and cell-invasion assay
Bania J, Zarczyńska A, Molenda J, Dabrowska A, Kosek-Paszkowska K, Wieckowska-Szakiel M, Rózalska B.
Language English Country Czech Republic
- MeSH
- Bacterial Proteins genetics MeSH
- Cytological Techniques MeSH
- DNA Fingerprinting methods MeSH
- Phylogeny MeSH
- Humans MeSH
- Listeria monocytogenes genetics isolation & purification classification MeSH
- Listeriosis microbiology MeSH
- Membrane Proteins genetics MeSH
- Cell Line, Tumor MeSH
- Polymerase Chain Reaction MeSH
- Food Microbiology MeSH
- Bacterial Typing Techniques methods MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
Analysis of actA gene sequence polymorphism has been shown to be an effective and relatively inexpensive method for subtyping Listeria monocytogenes isolates, allowing the division of the population of this species into two deeply separate lineages. This sequence-based method as well as PCR-mediated fingerprinting were applied here for the differentiation of 49 isolates of food and clinical origin. Correlation between these two typing approaches was high. Both methods divided the isolates into two lineages, designated I (33 isolates) and II (16 isolates). All the 33 lineage I isolates were assigned to the same, or closely related, six clusters by both typing methods. For the lineage II isolates, PCR fingerprinting was found to be more discriminatory. The isolates were characterized by cell invasion assay. All highly invasive isolates were assigned to lineage I, which constituted a heterogeneous group also containing low-invasive isolates. High-invasive isolates were not found in the genetically determined lineage II. A particular actA cluster, designated Ha, contained all the isolates showing the lowest invasiveness. A common trait of the isolates belonging to this cluster was the presence of a threonine-441 of the deduced ActA sequence instead of the alanine-441 present in the remaining isolates. Thirteen human isolates were classified to lineage I and five to lineage II. A PCR-based method can therefore differentiate L. monocytogenes isolates in accordance with the current phylogenetic model of the evolution of this species.
Lit.: 23
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- $a Department of Food Hygiene and Consumer Protection, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław
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- $a Lit.: 23
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- $a Analysis of actA gene sequence polymorphism has been shown to be an effective and relatively inexpensive method for subtyping Listeria monocytogenes isolates, allowing the division of the population of this species into two deeply separate lineages. This sequence-based method as well as PCR-mediated fingerprinting were applied here for the differentiation of 49 isolates of food and clinical origin. Correlation between these two typing approaches was high. Both methods divided the isolates into two lineages, designated I (33 isolates) and II (16 isolates). All the 33 lineage I isolates were assigned to the same, or closely related, six clusters by both typing methods. For the lineage II isolates, PCR fingerprinting was found to be more discriminatory. The isolates were characterized by cell invasion assay. All highly invasive isolates were assigned to lineage I, which constituted a heterogeneous group also containing low-invasive isolates. High-invasive isolates were not found in the genetically determined lineage II. A particular actA cluster, designated Ha, contained all the isolates showing the lowest invasiveness. A common trait of the isolates belonging to this cluster was the presence of a threonine-441 of the deduced ActA sequence instead of the alanine-441 present in the remaining isolates. Thirteen human isolates were classified to lineage I and five to lineage II. A PCR-based method can therefore differentiate L. monocytogenes isolates in accordance with the current phylogenetic model of the evolution of this species.
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