This process was carefully observed to prevent any loss of potentially discriminatory peaks at both ends of the derivative curves. To prevent excessive simplification and loss of informative data, smoothing was performed only if it undoubtedly resulted in a distinct amelioration of peaks’ discrimination. Electrophoresis and analysis of banding patterns After melting analysis was performed, each sample was also subjected to gel electrophoresis in 2% agarose gel at 5 V/cm for 3 hours. The gels were stained by ethidium bromide
MLN2238 in vitro added into them during preparation at the final concentration of 1 μg/ml and resulting banding patterns were photographed. Comparison of fingerprints was performed using GelCompar II software (Applied Maths, Sint-Martens-Latem, Belgium) applying the Jaccard coefficient at 1.5% positioning tolerance. Dendrograms were constructed using the UPGMA algorithm. Acknowledgements Ministry of Health (NR8365-4/2005), Czech Republic, supported this work. Dr. Mine Yücesoy
from Dokuz Eylül University, Izmir, Turkey and Dr. Jozef Nosek from BI 2536 ic50 Comenius University in Bratislava, Slovakia kindly gifted check details some of the strains. Technical assistance of Mrs. Jana Novotna, Mrs. Jitka Cankarova, and Mrs. Ivana Dosedelova is highly acknowledged. Electronic supplementary material Additional file 1: Similarity coefficients. Listing of similarity coefficients obtained upon automated comparison of normalized melting curves within each species. (XLS 250 KB) Additional file 2: Dendrogram of RAPD fingerprints. Dendrogram based on RAPD fingerprints of all strains included in the study. Analysis of RAPD fingerprinting patterns always provided accurate identification except for 2 strains showing quite unique fingerprints (marked by arrows). For comparison of strain clustering between conventional RAPD and McRAPD, the strains of different species are color-coded by ground tint colors and their specific McRAPD genotypes
are indicated by different saturation of colors. In case a strain was not assigned to a specific McRAPD genotype, it is not color-coded. (PNG 3 MB) Additional file 3: Average derivative curves. Plots of average McRAPD first negative derivative curves of species and genotypes included in the study. (XLS 1 MB) Additional file 4: Listing of clinical isolates and reference strains included in this study. (PDF 93 KB) References 1. Hobson RP: The Interleukin-2 receptor global epidemiology of invasive Candida infections – is the tide turning? J Hosp Infect 2003, 55:159–168. quiz 233CrossRefPubMed 2. Warnock DW: Trends in the epidemiology of invasive fungal infections. Nippon Ishinkin Gakkai Zasshi 2007, 48:1–12.CrossRefPubMed 3. Krcmery V, Barnes AJ: Non- albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance. J Hosp Infect 2002, 50:243–260.CrossRefPubMed 4. Freydiere AM, Guinet R, Boiron P: Yeast identification in the clinical microbiology laboratory: phenotypical methods. Med Mycol 2001, 39:9–33.PubMed 5.