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Screening of C.?parapsilosis complex isolates was based on the evaluation of yeast micromorphology and biochemical tests using the ID32?C System (bioM��rieux, Marcy l��Etoile, France). For molecular identification, DNA was extracted from single colonies using the fast small-scale isolation procedure described previously [11]. The internal transcribed spacer sequence is a non-coding region this website of the conserved rRNA genes. Therefore, it presents a high rate of polymorphism and has been used as a tool for the identification of fungal species [12]. The internal transcribed spacer region was amplified and sequenced using the C.?parapsilosis-specific primers CparapITSout?F (5��-CCGTCGTGCTGGGGATAGAG-3��) and CparapITSout?R (5��-CATCGCACGGGATTCTCACC-3��). Nucleotide sequences were analysed using the Blastn program at the NCBI site (http://www.ncbi.nlm.nih.gov) for species identification. Antifungal susceptibility testing was performed using the CLSI broth microdilution method [13]. The following antifungal drugs were tested: amphotericin?B, 5-flucytosine, fluconazole, voriconazole, itraconazole and caspofungin. Although endpoint reading for caspofungin is not mentioned in the CLSI protocol, MIC endpoints were determined after 24?h, on the basis of a prominent www.selleckchem.com/products/lee011.html decrease in growth as compared with that of the drug-free growth control, as suggested by other authors [10]. Differences in antifungal susceptibility patterns among species were evaluated using ANOVA. After sequencing of amplicons from 141 isolates, 124 (88%) were identified as C.?parapsilosis, 13 (9%) as C.?orthopsilosis, and four (3%) as C.?metapsilosis. This finding is consistent with data previously published by Lockhart et?al. [10] after testing strains of the C.?parapsilosis complex from Latin America, and indicate that the prevalence of bloodstream infections caused by C.?orthopsilosis in that region (9%) is probably higher than that in Europe (3.5�C5.7%) and the USA (5%) [9,10]. We also found that the prevalence of C.?metapsilosis fungaemia in Brazil (3%) Otenabant was lower than that documented in Spain (6.9%) [9]. Table?1 shows the results of antifungal susceptibility tests. All isolates were equally susceptible to amphotericin?B, voriconazole, itraconazole and caspofungin. Despite the fact that MIC50 and MIC90 values for C.?metapsilosis isolates were considerably higher than those obtained for C.?parapsilosissensu?stricto and C.?orthopsilosis, no single isolate was considered to have Dose Dependent Susceptibility (DDS) and/or resistance to azoles. It is of note that Van Asbeck et?al. [14] also found that C.?metapsilosis strains were slightly less susceptible to fluconazole than were strains of the other species. Consistent with data previously reported by Tavanti et?al. [15], the 5-flucytosine MIC90 values obtained for our C.?orthopsilosis isolates were significantly higher than those for the C.?parapsilosis and C.?metapsilosis isolates (p?