albicans genotype A, (B) C. albicans genotype B, (C) C. albicans genotype C, (D) C. glabrata, (E) C. parapsilosis, (F) C. pelliculosa, (G) C. krusei genotype A, (H) C. krusei genotype
B, (I) C. krusei genotype C. Discussion Our results show that McRAPD Lonafarnib molecular weight offers a promising alternative to conventional phenotypic identification techniques. Surprisingly, simple visual inspection of derivative plots performed best among the approaches tested for interpretation of mere numerical McRAPD data. Its performance almost matched the performance of traditional RAPD fingerprinting. Compared to the automated processing developed and tested by ourselves, the time costs of simple visual evaluation were roughly equal when using a pre-made computer-aided plotting scheme. However, with a broader spectrum of yeast species and expanding database of McRAPD results, simple visual Selleck JSH-23 examination can become more time demanding and cumbersome. Therefore, it may be advantageous to test for a threshold score in automated matching which can guarantee flawless identification in the future. Then, the visual matching could be reserved for isolates failing to reach this score in automated matching. When looking at the accuracy of identification obtained in this study, this should be regarded critically in the light
of the fact that all of the evaluations were based on an artificially assembled set of strains. However, because this CYTH4 set comprised
almost 95% of species typically isolated from clinical samples, real performance in routine settings should not differ too much. An ongoing prospective study being performed by ourselves should prove this assumption. When evaluating the future potential of McRAPD, we should first consider the main advantages and disadvantages of the RAPD technique itself. It is well-known that RAPD is highly sensitive not only to minor inter-strain differences, but also to minor differences in experimental conditions, which can result in different profiles, compromising intra- and interlaboratory reproducibility. There are many factors that can influence the appearance or disappearance of bands, including Mg2+ concentration, primer/template concentration ratio, Taq polymerase concentration and source, the model of thermal cycler etc. [15–18]. Since we aimed to use RAPD/McRAPD primarily not for strain typing but for species identification purposes, we optimised the amplification conditions in favour of low interstrain variability. This efficiently prevented problems with intralaboratory Selleck ISRIB reproducibility, as clearly demonstrated in Figure 4 and discussed above. Of course, some problems may occur with interlaboratory reproducibility, mainly when using a different model of thermal cycler or a different Taq polymerase.