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. 2008 Feb;14(2):282-90.
doi: 10.3201/eid1402.070920.

Molecular typing of Australian Scedosporium isolates showing genetic variability and numerous S. aurantiacum

Laurence Delhaes  1 , Azian Harun, Sharon C A Chen, Quoc Nguyen, Monica Slavin, Christopher H Heath, Krystyna Maszewska, Catriona Halliday, Vincent Robert, Tania C Sorrell; Australian Scedosporium (AUSCEDO) Study Group; Wieland Meyer
Affiliations

Molecular typing of Australian Scedosporium isolates showing genetic variability and numerous S. aurantiacum

Laurence Delhaes et al. Emerg Infect Dis. 2008 Feb.

Erratum in

  • Emerg Infect Dis. 2008 Apr;14(4):696

Abstract

One hundred clinical isolates from a prospective nationwide study of scedosporiosis in Australia (2003-2005) and 46 additional isolates were genotyped by internal transcribed spacer-restriction fragment length polymorphism (ITS-RFLP) analysis, ITS sequencing, and M13 PCR fingerprinting. ITS-RFLP and PCR fingerprinting identified 3 distinct genetic groups. The first group corresponded to Scedosporium prolificans (n = 83), and the other 2 comprised isolates previously identified as S. apiospermum: one of these corresponded to S. apiospermum (n = 33) and the other to the newly described species S. aurantiacum (n = 30). Intraspecies variation was highest for S. apiospermum (58%), followed by S. prolificans (45%) and S. aurantiacum (28%) as determined by PCR fingerprinting. ITS sequence variation of 2.2% was observed among S. apiospermum isolates. No correlation was found between genotype of strains and their geographic origin, body site from which they were cultured, or colonization versus invasive disease. Twelve S. prolificans isolates from 2 suspected case clusters were examined by amplified fragment length polymorphism analysis. No specific clusters were confirmed.

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Figures

Figure 1
Figure 1
Internal transcribed spacer–restriction fragment length polymorphism (ITS-RFLP) patterns obtained by double digestion with the enzymes Sau96I and HhaI (A) and of the PCR fingerprinting profiles obtained with the microsatellite specific primer M13 (B) for Scedosporium prolificans: lane 1, WM 06.457; lane 2, WM 06.458; lane 3, WM 06.503; lane 4, WM 06.502; lane 5, WM 06.399; lane 6, WM 06.434. S. aurantiacum: lane 7, WM 06.495; lane 8, WM 06.496; lane 9, WM 06.386; lane 10, WM 06.385; lane 11, WM 06.482; lane 12, WM 06.390. S. apiospermum: lane 13, WM 06.475; lane 14, WM 06.474; lane 15, WM 06.472; lane 16, WM 06.471; lane 17, WM 06.424; lane 18, WM 06.443; lane M, 1-kb marker (GIBCO-BRL, Gaithersburg, MD, USA).
Figure 2
Figure 2
Rooted phylogram (outgroup Pseudallescheria africana CBS 311.72 and Petriella setifera CBS 164.74), showing the relationships among 11 selected strains representing each obtained internal transcribed spacer (ITS)–restriction fragment length polymorphism pattern and 4 reference strain sequences obtained from GenBank by using PAUP* version 4.06.10 (29).
Figure 3
Figure 3
Three-dimensional presentation of the principal coordinate analysis of the PCR fingerprinting data showing 3 distinct clusters which correspond to Scedosporium prolificans (black dots), S. aurantiacum (dark gray dots), and S. apiospermum (light gray dots), with S. apiospermum showing the highest genetic variation.
Figure 4
Figure 4
Three-dimensional presentation of the principal coordinate analysis of the combined M13 PCR fingerprinting, amplified fragment length polymorphism (AFLP) primers EcoRI-GT and MseI-GT and AFLP primers EcoRI-TG and MseI-CA data from the suspected Sydney and Melbourne case cluster isolates and 23 other Australian isolates. None of the investigated isolates showed any epidemiologic connection except 3 isolates obtained from the same patient (nos. 1, 73, 119). Blue dots, Melbourne outbreak isolates; pink dot, Melbourne-related isolate; red dots, Sydney outbreak isolates; green dots, Sydney-related isolates; yellow dots, unrelated Australian isolates.
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