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Journal of Clinical Microbiology, October 2008, p. 3412-3416, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00046-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Development and Validation of a Quantitative PCR Assay for Diagnosis of Scedosporiosis{triangledown}

Maria V. Castelli, Maria J. Buitrago, Leticia Bernal-Martinez, Alicia Gomez-Lopez, Juan L. Rodriguez-Tudela, and Manuel Cuenca-Estrella*

Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Spain

Received 9 January 2008/ Returned for modification 24 March 2008/ Accepted 29 July 2008

Scedosporium apiospermum and Scedosporium prolificans are fungal pathogens that can cause severe human infections, including disseminated mycosis in immunocompromised patients. Two real-time PCR (RT-PCR) assays for the diagnosis of these species were developed and validated for the classification of clinical strains and for the detection of DNA in clinical samples by use of a murine model of invasive infection. A total of 14 clinical strains and 141 samples, including blood, serum, and lung samples from infected CD1 mice, were analyzed. Each RT-PCR methodology used a species-specific molecular beacon probe targeting a highly conserved region of the fungal ribosomal DNA gene. Results showed 100% specificity and a detection limit of 10 fg of DNA for both assays. The sensitivities for the S. prolificans-specific PCR assay were 100% for cultured clinical strains, 95.5% for lung tissues, 85% for serum, and 83.3% for blood. For S. apiospermum, the sensitivities were 100% for clinical strains and 97.2%, 81.8%, and 54.5% for lung tissues, serum, and blood, respectively. Both techniques can be useful for clinical diagnosis, and further studies are warranted.

* Corresponding author. Mailing address: Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra Majadahonda-Pozuelo Km 2, 28220 Majadahonda, Spain. Phone: 34-91-8223661. Fax: 34-91-5097966. E-mail: mcuenca-estrella{at}isciii.es

{triangledown} Published ahead of print on 6 August 2008.

Journal of Clinical Microbiology, October 2008, p. 3412-3416, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.00046-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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