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

Younes Maaroufi,1 Corine Heymans,1 Jean-Marc De Bruyne,1 Valerie Duchateau,1 Hector Rodriguez-Villalobos,2 Michel Aoun,1 and Françoise Crokaert1*

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.

* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, Institut Jules Bordet, Rue Héger-Bordet 1, 1000 Brussels, Belgium. Phone: 322 541 3700. Fax: 322 541 3295. E-mail: francoise.crokaert{at}bordet.be.

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.



This article has been cited by other articles:

  • Muir, A., Jenkins, A. T. A., Forrest, G., Clarkson, J., Wheals, A. (2009). Rapid electrochemical identification of pathogenic Candida species. J Med Microbiol 58: 1182-1189 [Abstract] [Full Text]  
  • Wellinghausen, N., Siegel, D., Winter, J., Gebert, S. (2009). Rapid diagnosis of candidaemia by real-time PCR detection of Candida DNA in blood samples. J Med Microbiol 58: 1106-1111 [Abstract] [Full Text]  
  • 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]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»