This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Maaroufi, Y. Articles by Crokaert, F. Search for Related Content PubMed PubMed Citation Articles by Maaroufi, Y. Articles by Crokaert, F.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2003, p. 3293-3298, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3293-3298.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Candida albicans in Clinical Blood Samples by Using a TaqMan-Based PCR Assay

Department of Microbiology and Infectious Diseases, Institut Jules Bordet, 1000 Brussels,¹ Department of Microbiology, Hôpital Erasme, 1070 Brussels, Belgium²

Received 31 May 2002/ Returned for modification 24 July 2002/ Accepted 14 March 2003

We describe a rapid and reproducible PCR assay for quantitation of the Candida albicans ribosomal DNA (rDNA) in clinical blood samples based on the TaqMan principle (Applied Biosystems), in which a signal is generated by cleavage of a template-specific probe during amplification. We used two fluorogenic probes based on universal, fungus-specific primers, one for the detection of C. albicans species DNA and one for the detection of all Candida genus DNA. C. albicans blastoconidia mixed with whole blood in a titration experiment yielded a linear PCR signal over a range of 3 orders of magnitude. The TaqMan-based PCR assay for C. albicans exhibited a low limit of detection (5 CFU/ml of blood) and an excellent reproducibility (96 to 99%). While the C. albicans species-specific probe had 100% specificity for C. albicans, all Candida genus-specific probes cross-reacted with other organisms likely to coinfect patients with C. albicans infections. On the basis of these data, we determined the C. albicans loads with a species-specific probe from 122 blood samples from 61 hematology or oncology patients with clinically proven or suspected systemic Candida infections. Eleven positive samples exhibited a wide range of C. albicans loads, extending from 5 to 100,475 CFU/ml of blood. The sensitivity and specificity of the present assay were 100 and 97%, respectively, compared with the results of blood culture. These data indicate that the TaqMan-based PCR assay for quantitation of C. albicans with a species-specific probe provides an attractive alternative for the identification and quantitation of C. albicans rDNA in pure cultures and blood samples.

This article has been cited by other articles:

• Metwally, L., Fairley, D. J., Coyle, P. V., Hay, R. J., Hedderwick, S., McCloskey, B., O'Neill, H. J., Webb, C. H., Elbaz, W., McMullan, R. (2008). Improving molecular detection of Candida DNA in whole blood: comparison of seven fungal DNA extraction protocols using real-time PCR. J Med Microbiol 57: 296-303 [Abstract] [Full Text]  
• Zeng, X., Kong, F., Halliday, C., Chen, S., Lau, A., Playford, G., Sorrell, T. C. (2007). Reverse Line Blot Hybridization Assay for Identification of Medically Important Fungi from Culture and Clinical Specimens. J. Clin. Microbiol. 45: 2872-2880 [Abstract] [Full Text]  
• Metwally, L., Hogg, G., Coyle, P. V., Hay, R. J., Hedderwick, S., McCloskey, B., O'Neill, H. J., Ong, G. M., Thompson, G., Webb, C. H., McMullan, R. (2007). Rapid differentiation between fluconazole-sensitive and -resistant species of Candida directly from positive blood-culture bottles by real-time PCR. J Med Microbiol 56: 964-970 [Abstract] [Full Text]  
• Innings, A., Ullberg, M., Johansson, A., Rubin, C. J., Noreus, N., Isaksson, M., Herrmann, B. (2007). Multiplex Real-Time PCR Targeting the RNase P RNA Gene for Detection and Identification of Candida Species in Blood. J. Clin. Microbiol. 45: 874-880 [Abstract] [Full Text]  
• Fujita, S.-i., Takamura, T., Nagahara, M., Hashimoto, T. (2006). Evaluation of a newly developed down-flow immunoassay for detection of serum mannan antigens in patients with candidaemia.. J Med Microbiol 55: 537-543 [Abstract] [Full Text]  
• Playford, E. G., Kong, F., Sun, Y., Wang, H., Halliday, C., Sorrell, T. C. (2006). Simultaneous detection and identification of Candida, Aspergillus, and cryptococcus species by reverse line blot hybridization.. J. Clin. Microbiol. 44: 876-880 [Abstract] [Full Text]  
• Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
• Philip, A., Odabasi, Z., Matiuzzi, G., Paetznick, V. L., Tan, S.-W., Warmington, J., Rex, J. H., Ostrosky-Zeichner, L. (2005). Syscan3, a Kit for Detection of Anti-Candida Antibodies for Diagnosis of Invasive Candidiasis. J. Clin. Microbiol. 43: 4834-4835 [Abstract] [Full Text]  
• Maaroufi, Y., Ahariz, N., Husson, M., Crokaert, F. (2004). Comparison of Different Methods of Isolation of DNA of Commonly Encountered Candida Species and Its Quantitation by Using a Real-Time PCR-Based Assay. J. Clin. Microbiol. 42: 3159-3163 [Abstract] [Full Text]  
• Maaroufi, Y., De Bruyne, J.-M., Duchateau, V., Georgala, A., Crokaert, F. (2004). Early Detection and Identification of Commonly Encountered Candida Species from Simulated Blood Cultures by Using a Real-Time PCR-Based Assay. J. Mol. Diagn. 6: 108-114 [Abstract] [Full Text]