Identification of invasive fungal diseases in immunocompromised patients by combining an Aspergillus specific PCR with a multifungal DNA-microarray from primary clinical samples.
The increasing incidence of invasive fungal diseases (IFD), most of all invasive aspergillosis (IA) in immunocompromised patients emphasises the need to improve the diagnostic tools for detection of fungal pathogens. We investigated the diagnostic performance of a multifungal DNA-microarray detecting 15 different fungi [Aspergillus, Candida, Fusarium, Mucor, Rhizopus, Scedosporium and Trichosporon species (spp.)] in addition to an Aspergillus specific polymerase chain reaction (PCR) assay. Biopsies, bronchoalveolar lavage and peripheral blood samples of 133 immunocompromised patients (pts) were investigated by a multifungal DNA-microarray as well as a nested Aspergillus specific PCR assay. Patients had proven (n = 18), probable (n = 29), possible (n = 48) and no IFD (n = 38) and were mostly under antifungal therapy at the time of sampling. The results were compared to culture, histopathology, imaging and serology, respectively. For the non-Aspergillus IFD the microarray analysis yielded in all samples a sensitivity of 64% and a specificity of 80%. Best results for the detection of all IFD were achieved by combining DNA-microarray and Aspergillus specific PCR in biopsy samples (sensitivity 79%; specificity 71%). The molecular assays in combination identify genomic DNA of fungal pathogens and may improve identification of causative pathogens of IFD and help overcoming the diagnostic uncertainty of culture and/or histopathology findings, even during antifungal therapy.
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
T
T
Boch
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
M
M
Reinwald
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
P
P
Postina
Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany.|Clinical Trials Centre Cologne, ZKS Köln, BMBF 01KN1106, Centre for Integrated Oncology CIO Köln Bonn, Cologne, Germany.|Cologne Excellence Cluster on Cellular Stress Response in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.
O A
OA
Cornely
Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany.|German Centre for Infection Research, Partner site Bonn-Cologne, Cologne, Germany.
J J
JJ
Vehreschild
Department of Diagnostic and Interventional Radiology with Nuclear Medicine, Thoraxklinik at Heidelberg University Hospital, Heidelberg, Germany.
C P
CP
Heußel
Department II of Internal Medicine, Würzburg University Hospital, Würzburg, Germany.
W J
WJ
Heinz
Section of Infectious Diseases and Tropical Medicine and Division of Pulmonology, Medical University of Graz, Graz, Austria.
M
M
Hoenigl
Section of Infectious Diseases and Tropical Medicine and Division of Pulmonology, Medical University of Graz, Graz, Austria.
S
S
Eigl
Pediatric Hematology and Oncology, Children's Hospital, University of Frankfurt, Frankfurt, Germany.
T
T
Lehrnbecher
Regensburg University Hospital, Regensburg, Germany.
J
J
Hahn
Ludwigshafen General Hospital, Ludwigshafen, Germany.
B
B
Claus
Department of Pediatrics, University Hospital Schleswig-Holstein, Campus Lübeck, Lübeck, Germany.
M
M
Lauten
Department V of Internal Medicine, University Hospital Heidelberg, Heidelberg, Germany.
G
G
Egerer
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
M C
MC
Müller
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
S
S
Will
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
N
N
Merker
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
W-K
WK
Hofmann
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
D
D
Buchheidt
Department of Hematology and Oncology, University Hospital Mannheim, University of Heidelberg, Mannheim, Germany.
B
B
Spiess
D000328:Adult; D000935:Antifungal Agents / Q000627:therapeutic use; D001228:Aspergillosis / Q000097:blood / Q000175:diagnosis* / Q000000981:diagnostic imaging; D001232:Aspergillus fumigatus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D001483:Base Sequence; D001707:Biopsy, Needle; D018893:Bronchoalveolar Lavage; D004271:DNA, Fungal / Q000302:isolation & purification; D005260:Female; D006801:Humans; D016867:Immunocompromised Host; D008297:Male; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011859:Radiography; D012680:Sensitivity and Specificity
D000935:Antifungal Agents; D004271:DNA, Fungal
DNA-microarray; clinical samples; invasive fungal disease; multiplex fungal PCR