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Rapid identification of Aspergillus fumigatus within the section Fumigati

DOI: 10.1186/1471-2180-11-82

Keywords: Aspergillus, azole resistance, electrophoretic profile, invasive aspergillosis, molecular identification, mould, multiplex PCR, rodlet A, β-tubulin

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Abstract:

A multiplex PCR was developed using prior information based on β-tubulin (βtub) and rodlet A (rodA) partial gene sequences. PCR amplification of βtub and rodA fragments resulted in a distinctive electrophoretic pattern in A. fumigatus and N. udagawae. The polymorphisms found in the smallest amplified sequence of βtub (153 bp) and rodA (103 bp) genes were then compared among and within species of this taxonomic section. βtub was able to differentiate among 13 individual species and two groups of species that included the pathogenic fungus A. lentulus. A more limited number of sequences were available for rodA; nevertheless, we were able to distinguish Aspergillus viridinutans, N. hiratsukae and N. udagawae.The assay described in the present study proved to be specific and highly reproducible, representing a fast and economic way of targeting molecular identification of the relevant mould, A. fumigatus, in clinical laboratories.Aspergillosis is the most common invasive mould disease worldwide. Recently, molecular techniques have been applied to fungal diagnosis and to the identification of species, and new fungal species that are morphologically similar to A. fumigatus have been described, authenticated and included in section Fumigati [1-3]. Therefore, this section now includes a few anamorphous Aspergillus species and teleomorphic species that are found in the genus Neosartorya [4]. The characteristics of the colonies on standard culture media are often similar to A. fumigatus, but conidia may be rather distinct. Neosartorya species produce heat-resistant ascospores [4].Misidentification of fungal species within the section Fumigati has been increasingly reported by clinical laboratories. Species, such as Aspergillus lentulus, Aspergillus viridinutans, Aspergillus fumigatiaffinis, Aspergillus fumisynnematus, Neosartorya pseudofischeri, Neosartorya hiratsukae and Neosartorya udagawae, are frequently reported as A. fumigatus [1,2,5,6]. Some of these species have been

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