title: Validation of the PCR-dHPLC method for rapid identification of Candida glabrata phylogenetically related species in different biological matrices.
abstract: Since two new species phylogenetically related to Candida glabrata with slightly different phenotypes and antifungal susceptibility profiles have been described, it seems to be necessary from clinical point of view, to develop a rapid and accurate identification system in order to distinguish between these three fungal species. We studied the performance of denaturing high performance liquid chromatography (dHPLC) as a faster (less than 7 min) and alternative novel technique for simultaneous analysis of Candida species in different biological matrices. The analyses show the good low limit of detection (LLOD) in all biological matrices studied (5.16-9.56 ngμL(-1), 4.14-4.70 ng μL(-1) and 3.99-4.66 ng μL(-1) for Candida bracarensis, Candida nivariensis and C. glabrata, respectively). 180 Candida isolates were analyzed in order to demonstrate the method suitability for screening analysis to identify C. glabrata and its cryptic species (C. bracarensis and C. nivariensis) in clinical routine.
authors.affiliation: Department of Clinical Microbiology and Infection Control, Basurto Hospital, Avenida Montevideo 18, 48013 Bilbao, Spain. otelleria001@ikasle.ehu.es
authors.forename: O
authors.initials: O
authors.lastname: Telleria
authors.forename: G
authors.initials: G
authors.lastname: Ezpeleta
authors.forename: O
authors.initials: O
authors.lastname: Herrero
authors.forename: I
authors.initials: I
authors.lastname: Miranda-Zapico
authors.forename: G
authors.initials: G
authors.lastname: Quindós
authors.forename: R
authors.initials: R
authors.lastname: Cisterna
mesh_terms: D001769:Blood / Q000382:microbiology; D002175:Candida / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D041221:Candida glabrata / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D004269:DNA, Bacterial / Q000737:chemistry; D021901:DNA, Intergenic / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D009691:Nucleic Acid Denaturation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D013183:Sputum / Q000382:microbiology; D014556:Urine / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; D021901:DNA, Intergenic