This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Schurko, A. M.
Right arrow Articles by Klassen, G. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Schurko, A. M.
Right arrow Articles by Klassen, G. R.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, June 2004, p. 2411-2418, Vol. 42, No. 6
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.6.2411-2418.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a Species-Specific Probe for Pythium insidiosum and the Diagnosis of Pythiosis

Andrew M. Schurko,1,{dagger} Leonel Mendoza,2 Arthur W. A. M. de Cock,3 James E. J. Bedard,1,{ddagger} and Glen R. Klassen1*

Department of Microbiology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada,1 Medical Technology Program, Department of Microbiology, Michigan State University, East Lansing, Michigan 48824-1031,2 Centraalbureau voor Schimmelcultures, NL-3508 AD Utrecht, The Netherlands3

Received 4 December 2003/ Returned for modification 12 February 2004/ Accepted 11 March 2004

Pythium insidiosum, the only species in the genus that infects mammals, is the etiological agent of pythiosis, a granulomatous disease characterized by cutaneous and subcutaneous lesions and vascular diseases. Accurate diagnosis of pythiosis and identification of its causal agent are often inconsistent with current immunological diagnostic methods. A species-specific DNA probe was constructed by using a 530-bp HinfI fragment from the ribosomal DNA intergenic spacer of P. insidiosum. When the probe was incubated with dot blots of genomic DNA from 104 Pythium species, it hybridized only to the DNA of P. insidiosum and P. destruens—two species that have been considered conspecific. The probe also hybridized to DNA from 22 P. insidiosum isolates in this study, regardless of their geographic origin or animal host. When tested against genomic DNA from other pathogenic organisms (Aspergillus fumigatus, Basidiobolus ranarum, Conidiobolus coronatus, Lagenidium giganteum, Paracoccidioides brasiliensis, and Prototheca wickerhamii), no cross-hybridization of the probe was detected. The specificity of the probe to hybridize to genomic DNA from all isolates of P. insidiosum and not cross-react with DNA from other Pythium species or pathogens that cause symptoms similar to pythiosis in their hosts makes it a powerful tool for the accurate diagnosis of pythiosis. In addition, the probe has the potential for pathological and environmental diagnostic applications.

* Corresponding author. Mailing address: Department of Microbiology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada. Phone: (204) 474-6303. Fax: (204) 474-7603. E-mail: glen_klassen{at}umanitoba.ca.

{dagger} Present address: Department of Biological Sciences, University of Iowa, Iowa City, IA 52242.

{ddagger} Present address: Department of Genome Science, Genome Research Institute, University of Cincinnati, Cincinnati, OH 45237.

Journal of Clinical Microbiology, June 2004, p. 2411-2418, Vol. 42, No. 6
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.6.2411-2418.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»