Development and Validation of a Multiplex PCR for Detection of Scedosporium spp. in Respiratory Tract Specimens from Patients with Cystic Fibrosis

Abstract

The emergence of Scedosporium infections in diverse groups of individuals, which are often treatment refractory, warrants timely and accurate laboratory diagnosis. Species-or group-specific primers based on internal transcribed spacer (ITS) sequence polymorphisms were designed for Scedosporium aurantiacum, Scedosporium dehoogii, Scedosporium prolificans, Pseudallescheria boydii species complex (former clade 5)/Pseudallescheria apiosperma (formerly classified as S. apiospermum sensu lato) and Pseudallescheria minutispora. Primers for S. aurantiacum, S. prolificans, and P. boydii species complex/P. apiosperma were incorporated into a multiplex PCR assay for the detection and identification of the three major clinically important Scedosporium species and validated using sputum specimens collected from patients seen at a major Australian cystic fibrosis clinic. The multiplex PCR assay showed 100% specificity in identifying the three major clinically relevant Scedosporium species from pure culture. When evaluated using DNA extracts from sputa, sensitivity and specificity of the multiplex PCR assay were 62.1% and 97.2%, respectively. This highly species-specific multiplex PCR assay offers a rapid and simple method of detection of the most clinically important Scedosporium species in respiratory tract specimens.

Detection, Identification, and Distribution of Fungi in Bronchoalveolar Lavage Specimens by Use of Multilocus PCR Coupled with Electrospray Ionization/Mass Spectrometry

“…However, it may take days or weeks to grow fungal pathogens by culture and an additional 1 to 2 days to identify these fungi by standard biochemical and phenotypic testing methods (4)(5)(6). Moreover, considerable experience is needed at the bench to identify mold-form fungi to the species level.…”

Purification and Characterization of a Mycelial Catalase from Scedosporium boydii, a Useful Tool for Specific Antibody Detection in Patients with Cystic Fibrosis

“…As for mycological examination, it requires skill and expertise and may lead to false-negative results for polymicrobial specimens like sputum samples because of the more rapid and more extensive growth of other molds frequently associated with this fungus, like Aspergillus fumigatus (19). Several molecular methods have been proposed for detection of the fungus from sputum samples (20)(21)(22)(23)(24), but as culture methods, they do not allow the differentiation between airway colonization and sensitization of the patient or respiratory infection in the CF context, which has important implications for patient management.…”

Rapid development of a mycotic aneurysm of the intracranial artery secondary to Scedosporium apiospermum sinusitis

“…Although the identification of S. apiospermum is generally based on mycology culture, the usefulness of several gene assays has been reported [10], owing to the delay in colony formation. In the present case, we demonstrated the usefulness of the molecular identification.…”