6533b861fe1ef96bd12c43b6

RESEARCH PRODUCT

Differentiation of Candida parapsilosis, C. orthopsilosis, and C. metapsilosis by specific PCR amplification of the RPS0 intron

Emilia CantónJavier PemánJosé M. García MartínezM. Micaela Gómez GarcíaM. Del Pilar VercherEulogio Valentín GómezLucas Del Castillo Agudo

subject

Microbiology (medical)Antifungal AgentsSequence analysisGenes FungalMolecular Sequence DataVirulenceMicrobial Sensitivity TestsBiologyCandida parapsilosisPolymerase Chain ReactionMicrobiologyMicrobiologylaw.inventionSpecies SpecificityDrug Resistance FungallawCloning MolecularDNA FungalMycological Typing TechniquesGenePolymerase chain reactionCandidaDNA PrimersGeneticsBase SequenceIntronFungal geneticsSequence Analysis DNAGeneral Medicinebiology.organism_classificationIntronsRandom Amplified Polymorphic DNA TechniqueRAPDInfectious Diseases

description

Although Candida parapsilosis is the most prevalent among the 3 species of the *psilosis group, studies applying DNA-based diagnostic techniques with isolates previously identified as C. parapsilosis have revealed that both C. orthopsilosis and C. metapsilosis account for 0-10% of all these isolates, depending on the geographical area. Differences in the degrees of antifungal susceptibility and virulence have been found, so a more precise identification is required. In a first approach, we reidentified 38 randomly chosen clinical isolates, previously identified as C. parapsilosis, using the RPO2 (CA2) RAPD marker. Among them, we reclassified 4 as C. metapsilosis and 5 as C. orthopsilosis. We previously developed a method to identify different pathogen yeast species, including C. parapsilosis, based on the amplification of the RPS0 gene intron. In this work, we extend this approach to the new *psilosis species by partially sequencing their RPS0 gene, including the intron sequence. Based on intron sequences, we designed specific primers capable of identifying C. orthopsilosis and C. metapsilosis species, and we reidentified species among the initial isolates. These new primers have allowed a specific and rapid identification of C. orthopsilosis and C. metapsilosis.

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