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Journal of Clinical Microbiology, August 1999, p. 2434-2438, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Specific Detection of Fusarium Species in Blood and Tissues by a PCR Technique

Francois-Xavier Hue,1,* Michel Huerre,2 Marie Ange Rouffault,1 and Claude de Bievre1

Laboratoire de Mycologie Médicale1 and Laboratoire d'Histopathologie,2 Institut Pasteur, 75724 Paris cedex 15, France

Received 14 October 1998/Returned for modification 1 February 1999/Accepted 30 April 1999

Fusarium species are opportunistic nosocomial pathogens that often cause fatal invasive mycoses. We designed a primer pair that amplifies by PCR a fragment of a gene coding for the rRNA of Fusarium species. The DNAs of the main Fusarium species and Neocosmospora vasinfecta but not the DNAs from 11 medically important fungi were amplified by these primers. The lower limit of detection of the PCR system was 10 fg of Fusarium solani DNA by ethidium bromide staining. To test the ability of this PCR system to detect Fusarium DNA in tissues, we developed a mouse model of disseminated fusariosis. Using the PCR, we detected Fusarium DNA in mouse tissues and in spiked human blood. Furthermore, F. solani, Fusarium moniliforme, and Fusarium oxysporum were testing by random amplified polymorphic DNA (RAPD) analysis. The bands produced by RAPD analysis were purified, cloned, and sequenced. The information was used to design primer pairs that selectively amplified one or several Fusarium species. The method developed may be useful for the rapid detection and identification of Fusarium species both from culture and from clinical samples.

* Corresponding author. Present address: Institut Pasteur, Morne Jolivière, BP 484, 97165 Pointe à Pitre cedex, Guadeloupe, France. Phone: 590.896940. Fax: 590896941. E-mail: cneyret{at}ipagua.gp.

Journal of Clinical Microbiology, August 1999, p. 2434-2438, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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Copyright © 1999 by the American Society for Microbiology. All rights reserved.