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Journal of Clinical Microbiology, April 1999, p. 925-930, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Serum Is More Suitable than Whole Blood for Diagnosis of Systemic Candidiasis by Nested PCR

M.-E. Bougnoux,1,* C. Dupont,1,2 J. Mateo,3 P. Saulnier,4 V. Faivre,3 D. Payen,3 and M.-H. Nicolas-Chanoine1

Departments of Microbiology1 and Internal Medicine,2 Hôpital Ambroise-Paré, Université Paris V, 92100 Boulogne-Billancourt, Department of Anesthesiology and Intensive Care, Hôpital Lariboisière, Université Paris VII, 75010 Paris,3 and Department of Microbiology, Institut Gustave Roussy, 94805 Villejuif,4 France

Received 3 November 1998/Returned for modification 7 December 1998/Accepted 13 January 1999

PCR assays for the diagnosis of systemic candidiasis can be performed either on serum or on whole blood, but results obtained with the two kinds of samples have never been formally compared. Thus we designed a nested PCR assay in which five specific inner pairs of primers were used to amplify specific targets on the rRNA genes of Candida albicans, C. tropicalis, C. parapsilosis, C. krusei, and C. glabrata. In vitro, the lower limit of detection of each nested PCR assay was 1 fg of purified DNA from the corresponding Candida species. In rabbits with candidemia of 120 minutes' duration following intravenous (i.v.) injection of 108 CFU of C. albicans, the sensitivities of the PCR in serum and whole blood were not significantly different (93 versus 86%). In other rabbits, injected with only 105 CFU of C. albicans, detection of candidemia by culture was possible for only 1 min, whereas DNA could be detected by PCR in whole blood and in serum for 15 and 150 min, respectively. PCR was more often positive in serum than in whole blood in 40 culture-negative samples (27 versus 7%; P < 0.05%). Lastly, experiments with rabbits injected i.v. with 20 or 200 µg of purified C. albicans DNA showed that PCRs were positive in serum from 30 to at least 120 min after injection, suggesting that the clearance of free DNA is slow. These results suggest that serum is the sample of choice, which should be used preferentially over whole blood for the diagnosis of systemic candidiasis by PCR.

* Corresponding author. Mailing address: Service de Microbiologie, Hôpital Ambroise-Paré, 9, avenue du Général de Gaulle, 92100 Boulogne-Billancourt, France. Phone: 33 (0) 1 49 09 55 45. Fax: 33 (0) 1 49 09 59 21. E-mail: marie-elisabeth.bougnoux{at}apr.ap-hop-paris.fr.

Journal of Clinical Microbiology, April 1999, p. 925-930, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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