title: Gene Scanning of an Internalin B Gene Fragment Using High-Resolution Melting Curve Analysis as a Tool for Rapid Typing of Listeria monocytogenes.
abstract: The ability to accurately track Listeria monocytogenes strains involved in outbreaks is essential for control and prevention of listeriosis. Because current typing techniques are time-consuming, cost-intensive, technically demanding, and difficult to standardize, we developed a rapid and cost-effective method for typing of L. monocytogenes. In all, 172 clinical L. monocytogenes isolates and 20 isolates from culture collections were typed by high-resolution melting (HRM) curve analysis of a specific locus of the internalin B gene (inlB). All obtained HRM curve profiles were verified by sequence analysis. The 192 tested L. monocytogenes isolates yielded 15 specific HRM curve profiles. Sequence analysis revealed that these 15 HRM curve profiles correspond to 18 distinct inlB sequence types. The HRM curve profiles obtained correlated with the five phylogenetic groups I.1, I.2, II.1, II.2, and III. Thus, HRM curve analysis constitutes an inexpensive assay and represents an improvement in typing relative to classical serotyping or multiplex PCR typing protocols. This method provides a rapid and powerful screening tool for simultaneous preliminary typing of up to 384 samples in approximately 2 hours.
authors.affiliation: Institute of Medical Microbiology and Hygiene, Austrian Agency for Health and Food Safety (AGES), Vienna, Austria.
authors.forename: Ariane T
authors.initials: AT
authors.lastname: Pietzka
authors.forename: Anna
authors.initials: A
authors.lastname: Stöger
authors.forename: Steliana
authors.initials: S
authors.lastname: Huhulescu
authors.forename: Franz
authors.initials: F
authors.lastname: Allerberger
authors.forename: Werner
authors.initials: W
authors.lastname: Ruppitsch
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D056726:Genetic Association Studies; D008089:Listeria monocytogenes / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D008565:Membrane Proteins / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012703:Serotyping
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; C095768:inlB protein, Listeria monocytogenes