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Journal of Clinical Microbiology, June 2009, p. 1818-1823, Vol. 47, No. 6
0095-1137/09/$08.00+0     doi:10.1128/JCM.02102-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Clinical Significance and Phylogenetic Relationship of Novel Australian Pneumocystis jirovecii Genotypes{triangledown}

Sebastiaan J. van Hal,1* Felix Gilgado,2 Tom Doyle,1 Joel Barratt,1 Damien Stark,1 Wieland Meyer,2 and Jock Harkness1

Department of Microbiology and Infectious Diseases, St. Vincents Hospital, Darlinghurst, Sydney, NSW, Australia,1 Molecular Mycology Research Laboratory, CIDM, University of Sydney Western Clinical School at Westmead Hospital, Westmead Millennium Institute, Westmead, NSW, Australia2

Received 2 November 2008/ Returned for modification 26 December 2008/ Accepted 28 March 2009

Pneumocystis jirovecii is an important opportunistic pathogen in immunocompromised patients. Molecular typing is employed to study this pathogen, as no culture system exists. No Australian P. jirovecii strains have been previously studied. Direct sequencing, targeting the internal transcribed spacer (ITS) regions of the nuclear rRNA operon, the mitochondrial large-subunit rRNA (mt LSU rRNA), and the dihydropteroate synthase (DHPS) gene, was performed on 68 Australian samples, collected between 2001 and 2007. Seven novel Australian ITS haplotypes (a composite of the ITS1 and ITS2 regions) were identified (SYD1m, SYD1g, Isyd2, Esyd3, Osyd4, Ag, and Hc). A dendrogram of published ITS haplotypes revealed that of the seven novel haplotypes, three (SYD1m, SYD1g, and Osyd4) are closely related to the haplotype Eg. Applying statistical parsimony, an Australian haplotype network was constructed which identified Eg as the ancestral haplotype, with two unresolved loops encountered. This suggests that the ITS lacks the resolution required for evolutionary analysis. Only two mt LSU rRNA genotypes were detected, with genotype 1 predominating. Mutant DHPS genotypes were present in 13% (8/60) of the samples. The novel haplotype Isyd2 was associated with less severe disease than the other Australian haplotypes. In contrast, patients with mutant DHPS genotypes were more likely to have severe disease, require invasive ventilation, and have a poor outcome than patients with wild-type DHPS genotypes. In conclusion, genetic clinical correlates continue to be found for Pneumocystis pneumonia; however, they remain controversial and warrant further study.

* Corresponding author. Mailing address: Department of Microbiology, St. Vincent's Hospital, Darlinghurst 2010, NSW Australia. Phone: 61 2 8382 9196. Fax: 61 2 8382 2989. E-mail: vanhal{at}gotalk.net.au

{triangledown} Published ahead of print on 15 April 2009.

Journal of Clinical Microbiology, June 2009, p. 1818-1823, Vol. 47, No. 6
0095-1137/09/$08.00+0     doi:10.1128/JCM.02102-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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