title: Development of species-specific primers for rapid identification of Debaryomyces hansenii.
abstract: In this work, we developed a specific PCR assay for Debaryomyces hansenii strains that uses a putative homologous PAD1 region (729 bp) present in this yeast species as a target. The amplification of this sequence with the D. hansenii specific primer pair (DhPADF/DhPADR) was found to be a rapid, specific and an affordable method enabling identification of D. hansenii from other yeast strains. Primers were tested in almost 100 strains, 49 strains from Type Culture Collection belonging to the genus Debaryomyces and to other yeast species commonly found in foods or related genera. These primers were able to discriminate between closely related species of Debaryomyces, such as Debaryomyces fabryi and Debaryomyces subglobosus, with a 100% detection rate for D. hansenii. Also, the method was tested in 45 strains from different foods. Results confirmed the specificity of the PCR method and detected two earlier misidentifications of D. hansenii strains obtained by RFLP analysis of the 5.8S ITS rDNA region. Subsequently we confirmed by sequencing the D1/D2 domain of 26S rDNA that these strains belonged to D. fabryi. We call attention in this work to the fact that the RFLPs of the 5.8S ITS rDNA profiles of D. hansenii, D. fabryi and D. subglobosus are the same and this technique will thus lead to incorrect identifications.
authors.affiliation: Departamento de Microbiología III, Facultad de Biología, Universidad Complutense de Madrid, C/ José Antonio Nováis, 2, 28040 Madrid, Spain; CEI Campus Moncloa, UCM-UPM, Madrid, Spain.
authors.forename: Petra
authors.initials: P
authors.lastname: Wrent
authors.affiliation: Departamento de Microbiología III, Facultad de Biología, Universidad Complutense de Madrid, C/ José Antonio Nováis, 2, 28040 Madrid, Spain; CEI Campus Moncloa, UCM-UPM, Madrid, Spain.
authors.forename: Eva-María
authors.initials: EM
authors.lastname: Rivas
authors.affiliation: Departamento de Microbiología III, Facultad de Biología, Universidad Complutense de Madrid, C/ José Antonio Nováis, 2, 28040 Madrid, Spain; CEI Campus Moncloa, UCM-UPM, Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Gil de Prado
authors.affiliation: Departamento de Microbiología III, Facultad de Biología, Universidad Complutense de Madrid, C/ José Antonio Nováis, 2, 28040 Madrid, Spain; CEI Campus Moncloa, UCM-UPM, Madrid, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: Peinado
authors.affiliation: Departamento de Microbiología III, Facultad de Biología, Universidad Complutense de Madrid, C/ José Antonio Nováis, 2, 28040 Madrid, Spain; CEI Campus Moncloa, UCM-UPM, Madrid, Spain. Electronic address: siloniz@bio.ucm.es.
authors.forename: María-Isabel
authors.initials: MI
authors.lastname: de Silóniz
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics; D005516:Food Microbiology / Q000379:methods*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D004718:Saccharomycetales / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004275:DNA, Ribosomal
keywords: Debaryomyces hansenii; Foods; PCR; Rapid-identification; Species-specific primers; Yeasts