Molecular Typing of Australian<i>Scedosporium</i>Isolates Showing Genetic Variability and Numerous<i>S. aurantiacum</i>

Delhaès, Laurence; Harun, Azian; Chen, Sharon C.‐A.; Nguyen, Quoc; Slavin, Monica A; Heath, Christopher H.; Maszewska, Krystyna; Halliday, Catriona; Robert, Vincent; Sorrell, Tania C.; Meyer, Wieland

doi:10.3201/eid1402.070920

Cited by 76 publications

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“…Access to DNA sequencing further enabled us to identify the causative pathogen as S. apiospermum sensu stricto. Recent genetic analyses have revealed P. boydii to be now separated from S. apiospermum as a distinct species and identified a new species of Scedosporium that would previously have been designated "S. apiospermum" (8,10,11). None of the isolates recovered from patients described previously (Table 1) were identified to the species level using molecular methods.…”

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“…Species identification of all four S. apiospermum and P. boydii isolates (taken from finger, paravertebral abscess, cutaneous wrist lesion, and aortic wall tissue samples) was performed by standard morphological methods (7) and confirmed by DNA sequencing of the internal transcribed spacer (ITS1/2) region of the fungal rRNA gene cluster (8,11). All isolates were identified as S. apiospermum, with 100% sequence similarity to the type strain of S. apiospermum sensu stricto (strain CBS 117407; GenBank accession number AJ 888416) (10,12).…”

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Fatal Mycotic Aneurysms Due to Scedosporium and Pseudallescheria Infection

et al. 2011

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Angioinvasive complications of Scedosporium infections are rare. We report two cases of mycotic aneurysm, following apparent localized infection, due to Scedosporium apiospermum and Pseudallescheria boydii. The thoracoabdominal aorta was affected in one patient, and cerebral vessels were affected in the other. Despite voriconazole therapy and surgical resection, the patients died. Previously reported cases are reviewed. CASE REPORTSPatient A. A 55-year-old male with diabetes mellitus who was the recipient of a kidney transplant from a living related donor presented in 2009 with a left index finger lesion, following a trivial gardening injury. Immunosuppressive treatment included mycophenolate mofetil (500 mg thrice daily), tacrolimus (3 mg twice daily), and prednisolone (10 mg daily), with a course of therapy with methylprednisolone (1 g daily for 3 days) in the preceding 6 months. A culture of material from the lesion grew Scedosporium apiospermum and Pseudallescheria boydii. The in vitro MIC of voriconazole was 1 g/ml (5). X rays of the hand showed no evidence of osteomyelitis, and there were no abnormalities upon chest X ray and cerebral computed tomography (CT) scanning. He received 3 months of oral voriconazole (200 mg twice daily after a loading dose of 6 mg/kg twice daily), with unchanged immunosuppression, with apparent complete clinical response.Nine months later, the patient developed progressive, severe flank and lower back pain. He was afebrile and his blood pressure was 190/100 mmHg. There was bilateral soft tissue tenderness of the mid-lumbar region but no vertebral or abdominal tenderness. A CT scan of the abdomen revealed nonspecific thickening of the aortic wall at the level of the third and fourth lumbar vertebra (LV3 to LV4). Magnetic resonance imaging (MRI) of the thoracolumbar-sacral vertebra showed destruction of LV4 without evidence of discitis between LV3 and LV4. A paravertebral abscess was observed extending from the 10th thoracic vertebra (TV) to the LV4 level (Fig. 1a), in addition to inflammatory aortitis with aneurysm formation of the aorta from the level of TV9 to LV4 (Fig. 1b).Empirical therapy with vancomycin, rifampin, and ciprofloxacin was commenced. Results from blood cultures were negative for bacteria and fungi. Drainage of the paravertebral abscess yielded 20 ml of purulent material; no organisms were seen on Gram or Ziehl-Nielsen staining. S. apiospermum and P. boydii were cultured after 14 days of incubation. A single, 1-by 2-cm subcutaneous nodule then appeared over the patient's right wrist. Histopathological examination (Gromori-Grocott and periodic acid-Schiff [PAS] staining) of the excised lesion revealed granulomatous inflammation and septate hyaline fungal hyphae; S. apiospermum and P. boydii were recovered after culturing. Treatment with voriconazole was reinitiated (6 mg/kg twice daily and then 4 mg/kg twice daily) in association with a reduction in the intensity of the immunosuppressive regimen. Voriconazole serum levels were checked regularly (trough levels w...

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Fatal Mycotic Aneurysms Due to Scedosporium and Pseudallescheria Infection

et al. 2011

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“…Although Scedosporium prolificans is represented in the Bruker library, only 1/4 isolates was identified to the species level. Scedosporiosis is the second most common nonAspergillus mold infection in Australia with 24% of infections due to S. aurantiacum (19,21,29). Adoption of our database in other Australian centers has the potential to remove reliance on molecular approaches to identify Scedosporium to the species level, overcoming the limitations of other dedicated Scedosporium databases that utilize different software, thus limiting their wider application (8,12,30).…”

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Performance of Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting

et al. 2016

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dWe developed an Australian database for the identification of Aspergillus, Scedosporium, and Fusarium species (n ‫؍‬ 28) by matrix-assisted laser desorption ionization؊time of flight mass spectrometry (MALDI-TOF MS). In a challenge against 117 isolates, species identification significantly improved when the in-house-built database was combined with the Bruker Filamentous Fungi Library compared with that for the Bruker library alone (Aspergillus, 93% versus 69%; Fusarium, 84% versus 42%; and Scedosporium, 94% versus 18%, respectively). Rapid, accurate mold identification is important due to the widening spectrum of pathogens and species-specific differences in antifungal susceptibility (1-3). Matrix-assisted laser desorption ionizationϪtime of flight mass spectrometry (MALDI-TOF MS) has proven useful, but mold identification remains challenged by the limited access to validated purpose-built databases that are necessary because of small species and strain representations in commercial libraries (4-16).Given the prior poor performance of the Bruker Filamentous Fungi Library v1.0 (Bruker Daltonics, Bremen, Germany) for mold identification using the manufacturer-recommended broth-based protein extraction methods (in our laboratory Ͼ50% of isolates were not identified; internal data) and because the geographic generalizability of in-house-built databases is not yet known, we hypothesized that a MS library of molds relevant to our region (17-21) will improve identification. Here, we constructed an in-house database containing 117 strains (see Table S1 in the supplemental material) covering 28 species of Aspergillus, Scedosporium, and Fusarium encountered in Australia. Challenge isolates (also n ϭ 117; 21 species) comprising 55 Aspergillus, 45 Fusarium, and 17 Scedosporium clinical strains (Table 1) were then used to assess the performance of the Bruker library alone versus that of the Bruker library supplemented with the in-house library for species identification.All isolates were identified using phenotypic methods (22) with definitive identification by DNA sequencing of the internal transcribed spacer (ITS) (all isolates), ␤-tubulin (Aspergillus and Scedosporium spp.), and partial elongation factor-1alpha (EF-1␣) (to identify Fusarium to the species complex level) gene regions (23)(24)(25)(26). Sequence data were analyzed against the Centraalbureau voor Schimmelkultures (http://www.cbs.knaw.nl/Collections /BioloMICSSequences.aspx?fileϭall), International Society for Human and Animal Mycology ITS (http://its.mycologylab.org/), and Fusarium-ID (http://www.fusariumdb.org/index.php) databases, and species were assigned using published criteria (27).Protein extraction for MALDI-TOF MS was performed as previously described (11). The Bruker bacterial test standard (Bruker Daltonics) was used for calibration and Aspergillus ustus CBS 261.67T scoring of Ն2.00 was required for quality extraction and spectra acceptability (11). The in-house database was constructed using published protocols (11, 28) with 20 to 25 quality spe...

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“…We choose the AFLP (amplified fragment length polymorphisms) technique, because it represents a highly discriminatory method at the intraspecific level and previously has been shown to have good discriminatory power for fungal strain differentiation (O'Donnell et al 2004, Delhaes et al 2008. In AFLP analyses, fragments are amplified from random locations throughout an organism's genome in a highly reproducible manner (Vos et al 1995, O'Donnell et al 2004, Delhaes et al 2008.…”

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“…In AFLP analyses, fragments are amplified from random locations throughout an organism's genome in a highly reproducible manner (Vos et al 1995, O'Donnell et al 2004, Delhaes et al 2008.…”

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Population structure and diversity of the pathogenic fungus Aspergillus fumigatus isolated from different sources and geographic origins

Duarte-Escalante

Zúñiga

Ramírez

et al. 2009

Mem. Inst. Oswaldo Cruz

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Molecular Typing of AustralianScedosporiumIsolates Showing Genetic Variability and NumerousS. aurantiacum

Abstract: Molecular typing showed genetic diversity, dismissed 2 suspected outbreaks of scedosporiosis, and identified multiple strains of the newly described species S. aurantiacum .

Cited by 76 publications

References 36 publications

Fatal Mycotic Aneurysms Due to Scedosporium and Pseudallescheria Infection

Fatal Mycotic Aneurysms Due to Scedosporium and Pseudallescheria Infection

Performance of Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting

Population structure and diversity of the pathogenic fungus Aspergillus fumigatus isolated from different sources and geographic origins

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