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#Azor phone 2 cases manual#
The sequences were aligned using the ClustalX (version 1.8) computer program ( 46), followed by manual adjustments with a text editor. proliferatum, respectively, since they are the only TUB sequences of the species included in the study that are available in the database (Table (Table1). We included five reference sequences retrieved from GenBank corresponding to F. We used the primers TUB-F ( 9) and T22 ( 33) with an annealing temperature of 55☌, and the PCR products were purified using a GFX PCR DNA kit (Pharmacia Biotech, Cerdanyola, Spain). For DNA extraction, amplification, and sequencing, we followed the procedures previously described by Gilgado et al. In addition, 19 reference strains from the Centraalbureau voor Schimmelcultures (CBS) and the Agricultural Research Service (ARS/NRRC) were used for a comparison.įor the phylogenetic analysis, we sequenced the TUB region of the β-tubulin gene, which has proven to be highly phylogenetically informative in different molecular studies of the genus Fusarium ( 33, 34, 35). Growth rates were obtained from colonies on PDA at 25☌ after 4 and 10 days of incubation in the dark.
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The microscopic features were examined by making direct wet mounts with lactic acid from cultures on PDA and OA after 7 to 10 days of incubation. Sporodochia can be detected by examining the cultures under a stereoscopic microscope, usually after 7 days of incubation. Briefly, to study the microscopic and colony features, the isolates were usually subcultured on potato-dextrose agar (PDA Difco Laboratories, Detroit, MI) and on oatmeal agar (OA 30 g oat flakes, 1 g MgSO 4♷H 2O, 1.5 g KH 2PO 4, 15 g agar, 1 liter tap water), incubated at 25☌ in the dark. Those isolates were identified following the conventional morphological criteria ( 25, 28, 30). Twenty-nine of these isolates, which were mainly from clinical sources, were sent to our laboratory from different clinical centers for identification purposes. We included 48 Fusarium isolates in the study (Table (Table1). Isolates and morphological identification. The objectives of our study were (i) to evaluate the correlation between the morphological and the molecular identification of less-frequent Fusarium species isolates received by our laboratory and (ii) to determine the antifungal susceptibilities of isolates representative of those less-common Fusarium species of clinical interest identified molecularly. In recent years, the in vitro antifungal susceptibilities of the most frequent species of Fusarium have been evaluated ( 1, 3, 4, 40, 47), but only a few isolates of the less-common species have been studied. However, several recent studies have demonstrated the usefulness of molecular methods for the identification of those Fusarium species that are difficult to distinguish morphologically ( 1, 4, 47). The identification of fusaria to the species level is not easy, and in numerous clinical cases the etiological agent is reported as being a Fusarium sp. verticillioides isolates were susceptible to posaconazole and terbinafine and Fusarium thapsinum isolates to terbinafine ( 4). However, some in vitro data concerning particular species seem to be very promising and deserve to be investigated clinically. Since the species of Fusarium are generally resistant to all the available antifungal drugs ( 40), it could be considered that speciation of Fusarium is necessary only for epidemiological purposes. The real incidence of these species is unknown since they are poorly known and laboratorians and clinical microbiologists are not generally aware of their possible presence in human infections. These species have been associated with different types of infection, in particular with keratomycoses and other ocular infections ( 10) and with disseminated infections in immunocompromised patients ( 2, 6, 17, 20, 23, 24, 26, 39, 41, 43, 44). Some of these species are Fusarium chlamydosporum, Fusarium dimerum, Fusarium incarnatum, and the following other species that are included into the Gibberella fujikuroi species complex: Fusarium napiforme, Fusarium nygamai, Fusarium proliferatum, and Fusarium sacchari ( 30, 31). The most frequent species causing fusariosis are Fusarium solani, Fusarium oxysporum, and Fusarium verticillioides ( 1, 16, 47), but several other species are also found to cause human infections, although less frequently.
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