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Journal of Clinical Microbiology, March 2006, p. 805-810, Vol. 44, No. 3
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.3.805-810.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genetic Identification of the Main Opportunistic Mucorales by PCR-Restriction Fragment Length Polymorphism

M. Machouart,1* J. Larché,2* K. Burton,1 J. Collomb,1 P. Maurer,2 A. Cintrat,1 M. F. Biava,1 S. Greciano,2 A. F. A. Kuijpers,3 N. Contet-Audonneau,1 G. S. de Hoog,3 A. Gérard,2 and B. Fortier1

Service de Parasitologie-Mycologie,1 Service de Réanimation Médicale, CHU Brabois, 54511 Vandoeuvre-Les-Nancy, France,2 Centraalbureau voor Schimmelcultures, P.O. Box 85167, 3508 AD Utrecht, The Netherlands3

Received 9 September 2005/ Returned for modification 20 October 2005/ Accepted 3 January 2006

Mucormycosis is a rare and opportunistic infection caused by fungi belonging to the order Mucorales. Recent reports have demonstrated an increasing incidence of mucormycosis, which is frequently lethal, especially in patients suffering from severe underlying conditions such as immunodeficiency. In addition, even though conventional mycology and histopathology assays allow for the identification of Mucorales, they often fail in offering a species-specific diagnosis. Due to the lack of other laboratory tests, a precise identification of these molds is thus notoriously difficult. In this study we aimed to develop a molecular biology tool to identify the main Mucorales involved in human pathology. A PCR strategy selectively amplifies genomic DNA from molds belonging to the genera Absidia, Mucor, Rhizopus, and Rhizomucor, excluding human DNA and DNA from other filamentous fungi and yeasts. A subsequent digestion step identified the Mucorales at genus and species level. This technique was validated using both fungal cultures and retrospective analyses of clinical samples. By enabling a rapid and precise identification of Mucorales strains in infected patients, this PCR-restriction fragment length polymorphism-based method should help clinicians to decide on the appropriate treatment, consequently decreasing the mortality of mucormycosis.

* Corresponding author. Mailing address for Marie Machouart: Service de Parasitologie-Mycologie, CHU Brabois, 54511 Vandoeuvre-Les-Nancy, France. Phone: 33 3 83 15 43 92. Fax: 33 3 83 15 43 86. E-mail: m.machouart{at}chu-nancy.fr. Mailing address for Jérôme Larché (reprint orders): Service de Réanimation Médicale, CHU Brabois, 54511 Vandoeuvre-Les-Nancy, France. Phone: 33 3 83 15 40 79. Fax: 33 3 83 15 42 20. E-mail: j.larche{at}chu-nancy.fr.

Journal of Clinical Microbiology, March 2006, p. 805-810, Vol. 44, No. 3
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.3.805-810.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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