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Journal of Clinical Microbiology, June 1999, p. 1871-1875, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

PCR-Restriction Enzyme Analysis for Detection of Candida DNA in Blood from Febrile Patients with Hematological Malignancies

Giulia Morace,1,* Livio Pagano,2 Maurizio Sanguinetti,1 Brunella Posteraro,1 Luca Mele,2 Francesco Equitani,2 Giuseppina D'Amore,1 Giuseppe Leone,2 and Giovanni Fadda1

Institutes of Microbiologia1 and Semeiotica Medica,2 Facoltà di Medicina e Chirurgia "A. Gemelli," Università Cattolica del Sacro Cuore, 1-00168 Rome, Italy

Received 10 August 1998/Returned for modification 2 November 1998/Accepted 1 March 1999

Blood samples were drawn daily from 72 patients who had hematological malignancies, neutropenia, and fever and who had failed to respond to broad-spectrum antibiotics. Each sample was used for conventional fungal blood cultures and for detection and identification of Candida DNA by a PCR method with subsequent restriction enzyme analysis (REA) recently developed in our laboratory. The PCR method was able to detect five CFU of Candida spp. per ml of blood, and subsequent REA of the amplicons allowed the identification of the Candida species most commonly implicated in cases of candidiasis. Thirty-one patients were PCR-REA positive, and four of these patients were also culture positive. The ultimate diagnosis for 13 of these patients and 1 patient who was PCR-REA negative was disseminated candidiasis (confirmed by clinical data, multiple cultures, histology, autopsy, and/or ultrasonographic evidence of hepatosplenic candidiasis). The molecular method is significantly more sensitive than conventional fungal blood cultures and has a high negative predictive value (97.5%) for the development of disseminated candidiasis in neutropenic patients.

* Corresponding author. Present address: Istituto di Microbiologia, Facoltà di Medicina e Chirurgia, Università degli Studi di Milano, Via Pascal 36, 20133 Milan, Italy. Phone: 39-2-26601215. Fax: 39-2-26601218. E-mail: morace{at}imiucca.csi.unimi.it.

Journal of Clinical Microbiology, June 1999, p. 1871-1875, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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