Skip to main content
Top
Published in: European Journal of Clinical Microbiology & Infectious Diseases 9/2003

01-09-2003 | Concise Article

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

Authors: A. Imhof, C. Schaer, G. Schoedon, D. J. Schaer, R. B. Walter, A. Schaffner, M. Schneemann

Published in: European Journal of Clinical Microbiology & Infectious Diseases | Issue 9/2003

Login to get access

Abstract

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.
Literature
1.
go back to reference 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 33:319–325PubMed 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 33:319–325PubMed
2.
go back to reference Duthie R, Denning DW (1995) Aspergillus fungemia: report of two cases and review. Clin Infect Dis 20:598–605PubMed Duthie R, Denning DW (1995) Aspergillus fungemia: report of two cases and review. Clin Infect Dis 20:598–605PubMed
3.
go back to reference Gottfredsson M, Cox GM, Perfect JR (1998) Molecular methods for epidemiological and diagnostic studies of fungal infections. Pathology 30:405–418PubMed Gottfredsson M, Cox GM, Perfect JR (1998) Molecular methods for epidemiological and diagnostic studies of fungal infections. Pathology 30:405–418PubMed
4.
go back to reference Walsh TJ, Chanock SJ (1998) Diagnosis of invasive fungal infections: advances in nonculture systems. Curr Clin Top Infect Dis 18:101–153PubMed Walsh TJ, Chanock SJ (1998) Diagnosis of invasive fungal infections: advances in nonculture systems. Curr Clin Top Infect Dis 18:101–153PubMed
5.
go back to reference Yeo SF, Wong B (2002) Current status of nonculture methods for diagnosis of invasive fungal infections. Clin Microbiol Rev 15:465–484CrossRefPubMed Yeo SF, Wong B (2002) Current status of nonculture methods for diagnosis of invasive fungal infections. Clin Microbiol Rev 15:465–484CrossRefPubMed
6.
go back to reference 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 38:4186–4192PubMed 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 38:4186–4192PubMed
7.
go back to reference 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 38:2902–2908PubMed 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 38:2902–2908PubMed
8.
go back to reference 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 39:1626–1629PubMed 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 39:1626–1629PubMed
9.
go back to reference 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 38:586–590PubMed 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 38:586–590PubMed
10.
go back to reference 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 20:744–745CrossRefPubMed 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 20:744–745CrossRefPubMed
11.
go back to reference Zinkernagel AS, Gmur R, Fenner L, Schaffner A, Schoedon G, Schneemann M (2003) Marginal and subgingival plaque—a natural habitat of Tropheryma whipplei? Infection 31:86–91PubMed Zinkernagel AS, Gmur R, Fenner L, Schaffner A, Schoedon G, Schneemann M (2003) Marginal and subgingival plaque—a natural habitat of Tropheryma whipplei? Infection 31:86–91PubMed
12.
go back to reference Olsen GJ, Woese CR (1993) Ribosomal RNA: a key to phylogeny. FASEB J 7:113–123PubMed Olsen GJ, Woese CR (1993) Ribosomal RNA: a key to phylogeny. FASEB J 7:113–123PubMed
13.
go back to reference Maleszka R, Clark-Walker GD (1993) Yeasts have a four-fold variation in ribosomal DNA copy number. Yeast 9:53–58PubMed Maleszka R, Clark-Walker GD (1993) Yeasts have a four-fold variation in ribosomal DNA copy number. Yeast 9:53–58PubMed
14.
go back to reference 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 47:811–820PubMed 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 47:811–820PubMed
15.
go back to reference Fredricks DN, Relman DA (1999) Application of polymerase chain reaction to the diagnosis of infectious diseases. Clin Infect Dis 29:475–486PubMed Fredricks DN, Relman DA (1999) Application of polymerase chain reaction to the diagnosis of infectious diseases. Clin Infect Dis 29:475–486PubMed
Metadata
Title
Rapid Detection of Pathogenic Fungi from Clinical Specimens Using LightCycler Real-Time Fluorescence PCR
Authors
A. Imhof
C. Schaer
G. Schoedon
D. J. Schaer
R. B. Walter
A. Schaffner
M. Schneemann
Publication date
01-09-2003
Publisher
Springer-Verlag
Published in
European Journal of Clinical Microbiology & Infectious Diseases / Issue 9/2003
Print ISSN: 0934-9723
Electronic ISSN: 1435-4373
DOI
https://doi.org/10.1007/s10096-003-0989-0

Other articles of this Issue 9/2003

European Journal of Clinical Microbiology & Infectious Diseases 9/2003 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine