When comparing operation costs of both procedures, our experience shows that McRAPD can be quite

competitive compared to ID 32C, however, market prices of materials and sets are always subject to change. Thus, it should be fair to say that both approaches are roughly comparable, McRAPD being more rapid with a potential Nirogacestat in vivo for future improvements. Since ID 32C offers the most extensive set of assimilation tests among commercially available yeast identification systems, it can be expected that other phenotyping approaches will show inferior performance. Thus, the need of special instrumentation and skills should be the only obstacle for general acceptance of McRAPD in routine diagnostic laboratories. Generally speaking, those laboratories being able to adopt McRAPD will be also able to adopt other genotyping techniques. Then,

such techniques, Multi Locus Sequence Typing (MLST) in particular, should be the main competitors of McRAPD. Although MLST is more demanding concerning instrumentation, ISRIB skills and labour, it has the advantage of unmatched interlaboratory reproducibility, enabling global epidemiology. However, it can hardly be expected that MLST can present an economically affordable alternative for routine identification and Oligomycin A manufacturer prospective epidemiological surveillance in near future. It can rather be expected that its use will be limited to retrospective epidemiological studies. Thus, McRAPD offers a promising choice for routine identification of pathogenic yeast species; Y-27632 in vivo in case of failure, it could be supplemented by other techniques, the best of which appears to be single-locus sequencing in our opinion. Conclusions 1. Crude colony

lysates provide an economical, rapid and reliable alternative to elaborate DNA extraction techniques for the purposes of McRAPD when performed by skilled personnel. 2. Our optimized McRAPD protocol shows excellent intralaboratory reproducibility and is able to delineate specific genotypes in some of the species studied. 3. Computer-aided visual matching of first derivative plots shows best performance among the approaches tested for interpretation of mere numerical McRAPD data. Its performance almost matched the performance of traditional RAPD fingerprinting and was comparable to the performance of the ID32C commercial system. 4. We believe that because of its advantages over conventional phenotypic identification approaches and competitive costs McRAPD can find its place in routine identification of medically important yeasts in advanced diagnostic laboratories being able to adopt the technique. It can also serve as a broad-range high-throughput technique for crude epidemiological surveillance. Methods Yeast strains The 9 yeast species most frequently isolated from clinical samples in our settings, namely representing 94.3% of yeast species isolated from patient samples at our department, were included into the study. Among these, 7 more common species, i.e. Candida albicans (56.2%), C.