Rapid Detection of Pathogenic Fungi from Clinical Specimens Using LightCycler Real-Time Fluorescence PCR

  • A. Imhof
  • C. Schaer
  • G. Schoedon
  • D. J. Schaer
  • R. B. Walter
  • A. Schaffner
  • M. SchneemannEmail author
Concise Article


In the study presented here a LightCycler real-time PCR system was used for the diagnosis of fungal infections from clinical tissue samples. Nine specimens were investigated from six patients with suspected or proven invasive fungal infections. Seven of nine samples were positive in a broad-range fungal PCR assay. In four samples, Aspergillus fumigatus was detected both by a species-specific hybridization assay as well as by sequencing of amplification products. In addition, the broad-range fungal PCR assay and PCR sequencing detected and identified, respectively, the following organisms in the specimens noted: Candida albicans in a culture-negative liver biopsy, Histoplasma capsulatum in a bone marrow sample, and Conidiobolus coronatus in a facial soft tissue specimen. Real-time PCR is a promising tool for the diagnosis of invasive fungal infections in human tissue samples and offers some advantages over culture methods, such as rapid analysis and increased sensitivity.


Aspergillus Fumigatus Invasive Fungal Infection Human Tissue Sample Histoplasma Capsulatum Facial Soft Tissue 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Haynes KA, Westerneng TJ, Fell JW, Moens W (1995) Rapid detection and identification of pathogenic fungi by polymerase chain reaction amplification of large subunit ribosomal DNA. J Med Vet Mycol 33:319–325PubMedGoogle Scholar
  2. 2.
    Duthie R, Denning DW (1995) Aspergillus fungemia: report of two cases and review. Clin Infect Dis 20:598–605PubMedGoogle Scholar
  3. 3.
    Gottfredsson M, Cox GM, Perfect JR (1998) Molecular methods for epidemiological and diagnostic studies of fungal infections. Pathology 30:405–418PubMedGoogle Scholar
  4. 4.
    Walsh TJ, Chanock SJ (1998) Diagnosis of invasive fungal infections: advances in nonculture systems. Curr Clin Top Infect Dis 18:101–153PubMedGoogle Scholar
  5. 5.
    Yeo SF, Wong B (2002) Current status of nonculture methods for diagnosis of invasive fungal infections. Clin Microbiol Rev 15:465–484CrossRefPubMedGoogle Scholar
  6. 6.
    Hendolin PH, Paulin L, Koukila-Kahkola P, Anttila VJ, Malmberg H, Richardson M, Ylikoski J (2000) Panfungal PCR and multiplex liquid hybridization for detection of fungi in tissue specimens. J Clin Microbiol 38:4186–4192PubMedGoogle Scholar
  7. 7.
    Jaeger EE, Carroll NM, Choudhury S, Dunlop AA, Towler HM, Matheson MM, Adamson P, Okhravi N, Lightman S (2000) Rapid detection and identification of Candida, Aspergillus, and Fusarium species in ocular samples using nested PCR. J Clin Microbiol 38:2902–2908PubMedGoogle Scholar
  8. 8.
    Loeffler J, Hebart H, Cox P, Flues N, Schumacher U, Einsele H (2001) Nucleic acid sequence-based amplification of Aspergillus RNA in blood samples. J Clin Microbiol 39:1626–1629PubMedGoogle Scholar
  9. 9.
    Loeffler J, Henke N, Hebart H, Schmidt D, Hagmeyer L, Schumacher U, Einsele H (2000) Quantification of fungal DNA by using fluorescence resonance energy transfer and the light cycler system. J Clin Microbiol 38:586–590PubMedGoogle Scholar
  10. 10.
    Rickerts V, Loeffler J, Bцhme A, Einsele H, Just-Nübling G (2001) Diagnosis of disseminated zygomycosis using a polymerase chain reaction assay. Eur J Clin Microbiol Infect Dis 20:744–745CrossRefPubMedGoogle Scholar
  11. 11.
    Zinkernagel AS, Gmur R, Fenner L, Schaffner A, Schoedon G, Schneemann M (2003) Marginal and subgingival plaque—a natural habitat of Tropheryma whipplei? Infection 31:86–91PubMedGoogle Scholar
  12. 12.
    Olsen GJ, Woese CR (1993) Ribosomal RNA: a key to phylogeny. FASEB J 7:113–123PubMedGoogle Scholar
  13. 13.
    Maleszka R, Clark-Walker GD (1993) Yeasts have a four-fold variation in ribosomal DNA copy number. Yeast 9:53–58PubMedGoogle Scholar
  14. 14.
    Kappe R, Okeke CN, Fauser C, Maiwald M, Sonntag HG (1998) Molecular probes for the detection of pathogenic fungi in the presence of human tissue. J Med Microbiol 47:811–820PubMedGoogle Scholar
  15. 15.
    Fredricks DN, Relman DA (1999) Application of polymerase chain reaction to the diagnosis of infectious diseases. Clin Infect Dis 29:475–486PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • A. Imhof
    • 1
  • C. Schaer
    • 3
  • G. Schoedon
    • 3
  • D. J. Schaer
    • 4
  • R. B. Walter
    • 2
  • A. Schaffner
    • 3
  • M. Schneemann
    • 3
    Email author
  1. 1.Program in Infectious Disease, Clinical Research DivisionFred Hutchinson Cancer Research CenterSeattleUSA
  2. 2.Pediatric Oncology, Clinical Research DivisionFred Hutchinson Cancer Research CenterSeattleUSA
  3. 3.Department of Medicine, Clinical Mycology, Medical Clinic BUniversity Hospital ZurichZurichSwitzerland
  4. 4.Department of AnesthesiologyUniversity Hospital ZurichZurichSwitzerland

Personalised recommendations