title: High resolution melting analysis (HRM) as a new tool for the identification of species belonging to the Lactobacillus casei group and comparison with species-specific PCRs and multiplex PCR.
abstract: The correct identification and characterisation of bacteria is essential for several reasons: the classification of lactic acid bacteria (LAB) has changed significantly over the years, and it is important to distinguish and define them correctly, according to the current nomenclature, avoiding problems in the interpretation of literature, as well as mislabelling when probiotic are used in food products. In this study, species-specific PCR and HRM (high-resolution melting) analysis were developed to identify strains belonging to the Lactobacillus casei group and to classify them into L. casei, Lactobacillus paracasei and Lactobacillus rhamnosus. HRM analysis confirmed to be a potent, simple, fast and economic tool for microbial identification. In particular, 201 strains, collected from International collections and attributed to the L. casei group, were examined using these techniques and the results were compared with consolidated molecular methods, already published. Seven of the tested strains don't belong to the L. casei group. Among the remaining 194 strains, 6 showed inconsistent results, leaving identification undetermined. All the applied techniques were congruent for the identification of the vast majority of the tested strains (188). Notably, for 46 of the strains, the identification differed from the previous attribution.
authors.affiliation: Dipartimento di Scienze degli Alimenti, Università degli Studi di Udine, Udine, Italy. Electronic address: lucilla.iacumin@uniud.it.
authors.forename: Lucilla
authors.initials: L
authors.lastname: Iacumin
authors.affiliation: Dipartimento di Scienze degli Alimenti, Università degli Studi di Udine, Udine, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Ginaldi
authors.affiliation: Dipartimento di Scienze degli Alimenti, Università degli Studi di Udine, Udine, Italy.
authors.forename: Marisa
authors.initials: M
authors.lastname: Manzano
authors.affiliation: Dipartimento di Scienze degli Alimenti, Università degli Studi di Udine, Udine, Italy.
authors.forename: Veronica
authors.initials: V
authors.lastname: Anastasi
authors.affiliation: Istituto di Scienze dell'Alimentazione-CNR, Avellino, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Reale
authors.affiliation: Istituto di Scienze dell'Alimentazione-CNR, Avellino, Italy.
authors.forename: Teresa
authors.initials: T
authors.lastname: Zotta
authors.affiliation: Dipartimento di Agricoltura, Ambiente e Alimenti Università degli Studi del Molise, Campobasso, Italy.
authors.forename: Franca
authors.initials: F
authors.lastname: Rossi
authors.affiliation: Istituto di Scienze dell'Alimentazione-CNR, Avellino, Italy; Dipartimento di Agricoltura, Ambiente e Alimenti Università degli Studi del Molise, Campobasso, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Coppola
authors.affiliation: Dipartimento di Scienze degli Alimenti, Università degli Studi di Udine, Udine, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Comi
mesh_terms: D004269:DNA, Bacterial / Q000737:chemistry* / Q000235:genetics; D007780:Lacticaseibacillus casei / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D044366:Transition Temperature
chemical_list: D004269:DNA, Bacterial
keywords: High resolution melting analysis; Identification methods; Lactobacillus casei group; Multiplex PCR; Species-specific PCR