title: Real-time PCR and quantitative culture for monitoring of experimental Aspergillus fumigatus intracranial infection in neutropenic mice.
abstract: The central nervous system (CNS) is the most common site of dissemination during Aspergillus infection. PCR has the potential to facilitate early diagnosis of CNS aspergillosis, which could assist in reducing disease mortality. In two experiments, neutropenic CD-1 male mice were infected intracranially with 5×10⁶ conidia of Aspergillus fumigatus. At time points up to 120 h after infection, mice were euthanized and samples of blood, brain, spinal cord and cerebrospinal fluid (CSF) were taken. The brain fungal burden was determined by quantitative culture, and fungal DNA was detected by quantitative PCR. Plating for A. fumigatus from the brain confirmed that all mice had burdens of log₁₀>3 from 4 to 120 h after infection. A. fumigatus DNA was detected in blood (88 %), brain (96 %), CSF (52 %) and spinal cord (92 %) samples. The brain and spinal cord contained the highest concentrations of fungal DNA. Adapting the extraction protocol to maximize yield from small sample volumes (10 µl CSF or 200 µl blood) allowed PCR detection of A. fumigatus in infected mice, suggesting the use of CSF and blood as diagnostic clinical samples for CNS aspergillosis.
authors: Department of Clinical Microbiology, Sir Patrick Dun Research Laboratory, School of Medicine, Trinity College Dublin, Dublin 8, Ireland.
authors: C Oliver
authors: CO
authors: Morton
authors: California Institute for Medical Research, San Jose, CA 95128, USA.|Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University, Stanford, CA 94305, USA.|Department of Medicine, Division of Infectious Diseases, Santa Clara Valley Medical Center, San Jose, CA 95128-2699, USA.
authors: Karl V
authors: KV
authors: Clemons
authors: Universität Wuerzburg, Medizinische Klinik & Poliklinik II, Wuerzburg, Germany.
authors: Jan
authors: J
authors: Springer
authors: Pathologisches Institut der Universität Wuerzburg, Wuerzburg, Germany.
authors: Justus G
authors: JG
authors: Mueller
authors: Department of Clinical Microbiology, Sir Patrick Dun Research Laboratory, School of Medicine, Trinity College Dublin, Dublin 8, Ireland.
authors: Thomas R
authors: TR
authors: Rogers
authors: Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University, Stanford, CA 94305, USA.|Department of Medicine, Division of Infectious Diseases, Santa Clara Valley Medical Center, San Jose, CA 95128-2699, USA.|California Institute for Medical Research, San Jose, CA 95128, USA.
authors: David A
authors: DA
authors: Stevens
authors: Friedrich-Schiller-Universität Jena, Septomics Research Centre, Leibniz Institute for Natural Products Research and Infection Biology - Hans Knöll Institute, Jena, Germany.
authors: Oliver
authors: O
authors: Kurzai
authors: Universität Wuerzburg, Medizinische Klinik & Poliklinik II, Wuerzburg, Germany.
authors: Hermann
authors: H
authors: Einsele
authors: Universität Wuerzburg, Medizinische Klinik & Poliklinik II, Wuerzburg, Germany.
authors: Juergen
authors: J
authors: Loeffler
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001921:Brain / Q000382:microbiology* / Q000473:pathology; D004271:DNA, Fungal / Q000134:cerebrospinal fluid; D005690:Galactose / Q000031:analogs & derivatives; D008297:Male; D008351:Mannans / Q000378:metabolism; D051379:Mice; D020953:Neuroaspergillosis / Q000382:microbiology*; D009503:Neutropenia / Q000150:complications*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013234:Stem Cells
chemical_list: D004271:DNA, Fungal; D008351:Mannans; C012990:galactomannan; D005690:Galactose