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Clinical and Vaccine Immunology, May 2009, p. 756-764, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00061-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Tracking the Emerging Human Pathogen Pseudallescheria boydii by Using Highly Specific Monoclonal Antibodies {triangledown}

Christopher R. Thornton*

Hybridoma Laboratory, School of Biosciences, Geoffrey Pope Building, University of Exeter, Stocker Road, Exeter, Devon EX4 4QD, United Kingdom

Received 10 February 2009/ Returned for modification 11 March 2009/ Accepted 20 March 2009

Pseudallescheria boydii has long been known to cause white grain mycetoma in immunocompetent humans, but it has recently emerged as an opportunistic pathogen of humans, causing potentially fatal invasive infections in immunocompromised individuals and evacuees of natural disasters, such as tsunamis and hurricanes. The diagnosis of P. boydii is problematic since it exhibits morphological characteristics similar to those of other hyaline fungi that cause infectious diseases, such as Aspergillus fumigatus and Scedosporium prolificans. This paper describes the development of immunoglobulin M (IgM) and IgG1 {kappa}-light chain monoclonal antibodies (MAbs) specific to P. boydii and certain closely related fungi. The MAbs bind to an immunodominant carbohydrate epitope on an extracellular 120-kDa antigen present in the spore and hyphal cell walls of P. boydii and Scedosporium apiospermum. The MAbs do not react with S. prolificans, Scedosporium dehoogii, or a large number of clinically relevant fungi, including A. fumigatus, Candida albicans, Cryptococcus neoformans, Fusarium solani, and Rhizopus oryzae. The MAbs were used in immunofluorescence and double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) to accurately differentiate P. boydii from other infectious fungi and to track the pathogen in environmental samples. Specificity of the DAS-ELISA was confirmed by sequencing of the internally transcribed spacer 1 (ITS1)-5.8S-ITS2 rRNA-encoding regions of environmental isolates.

* Mailing address: Hybridoma Laboratory, School of Biosciences, Geoffrey Pope Building, University of Exeter, Stocker Road, Exeter, Devon EX4 4QD, United Kingdom. Phone: 44 (0)1392 263776. Fax: 44 (0)1392 263434. E-mail: C.R.Thornton{at}ex.ac.uk

{triangledown} Published ahead of print on 25 March 2009.

Clinical and Vaccine Immunology, May 2009, p. 756-764, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00061-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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