fumigatiaffinis and A. lentulus [7, 8], whereas A. fumigatus is usually susceptible to the antifungals that are available for clinical treatment [19, 20]. Few clinical cases of invasive aspergillosis have been selleck chemicals reported in which the antifungal treatment was repeatedly modified until the correct identification of the fungal agent and the administration of the appropriate antifungal treatment [17, 18]. Considering that A. fumigatus may represent a considerable part of all clinical cases of aspergillosis, molecular characterization is essential for the correct identification of species within the section Fumigati. In this study, we developed a multiplex PCR strategy that was able
to differentiate A. fumigatus from all the other related species within the section Fumigati. Birinapant order We could not test all of the species of section Fumigati, as some of them are extremely rare. However, we believe that the present multiplex PCR can be widely used, as A. lentulus is more closely related to A. fumigatus than most species in section Fumigati (e.g. A. viridinutans) [4, 5], and a distinct electrophoretic profile was observed with two strains
of this species. It is expected that other species of section Fumigati that are genetically distant from A. fumigatus can be distinguished by employing TPX-0005 this multiplex PCR (see additional file 2 in supplemental data). A simple electrophoresis profile after PCR amplification clearly separates two species, A. fumigatus and N. udagawae, from a second group of fungal isolates of section Fumigati. This method is furthermore amenable to automation. Compared to previously described methodologies for A. fumigatus identification within its section [10–13], the proposed method facilitates the molecular recognition of this species by employing a single multiplex PCR and avoiding the need for restriction enzymes and specialized 2-hydroxyphytanoyl-CoA lyase equipment. This approach is cheap and simple and would be very useful
in clinical labs that routinely screen and perform the molecular identification of several mould isolates. The proposed new assay proved to be specific and highly reproducible for targeting A. fumigatus within the section Fumigati and outside this section. A list of fungal species related to A. fumigatus could be identified by sequencing partial regions of βtub and rodA. A group of 14 unique species and two groups of species of section Fumigati were distinguished by point mutations in βtub and rodA. This work presents the first record of polymorphic sites available for the rapid identification of species within the section Fumigati following the analysis of more than 450 βtub and rodA sequences. This list represents a practical guide for the molecular recognition of rare fungal species, and it can certainly be expanded in the near future when more sequences of βtub and rodA are available.