Document Detail


Bioluminescent Aspergillus fumigatus, a new tool for drug efficiency testing and in vivo monitoring of invasive aspergillosis.
MedLine Citation:
PMID:  18820063     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Aspergillus fumigatus is the main cause of invasive aspergillosis in immunocompromised patients, and only a limited number of drugs for treatment are available. A screening method for new antifungal compounds is urgently required, preferably an approach suitable for in vitro and in vivo studies. Bioluminescence imaging is a powerful tool to study the temporal and spatial resolutions of the infection and the effectiveness of antifungal drugs. Here, we describe the construction of a bioluminescent A. fumigatus strain by fusing the promoter of the glyceraldehyde-3-phosphate dehydrogenase gene from A. fumigatus with the luciferase gene from Photinus pyralis to control the expression of the bioluminescent reporter. A. fumigatus transformed with this construct revealed high bioluminescence under all tested growth conditions. Furthermore, light emission correlated with the number of conidia used for inoculation and with the biomass formed after different incubation times. The bioluminescent strains were suitable to study the effectiveness of antifungals in vitro by several independent methods, including the determination of light emission with a microplate reader and the direct visualization of light emission with an IVIS 100 system. Moreover, when glucocorticoid-treated immunosuppressed mice were infected with a bioluminescent strain, light emission was detected from infected lungs, allowing the visualization of the progression of invasive aspergillosis. Therefore, this new bioluminescence tool is suitable to study the in vitro effectiveness of drugs and the disease development, localization, and burden of fungi within tissues and may also provide a powerful tool to study the effectiveness of antifungals in vivo.
Authors:
Matthias Brock; Grégory Jouvion; Sabrina Droin-Bergère; Olivier Dussurget; Marie-Anne Nicola; Oumaïma Ibrahim-Granet
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-09-26
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  74     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2008 Nov 
Date Detail:
Created Date:  2008-11-12     Completed Date:  2008-12-11     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  7023-35     Citation Subset:  IM    
Affiliation:
Leibniz Institute for Natural Product Research and Infection Biology, Hans Knoell Institute, Junior Research Group Microbial Biochemistry and Physiology, Beutenbergstr. 11a, 07745 Jena, Germany. Matthias.brock@hki-jena.de
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AM999768;  CAQ53727
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MeSH Terms
Descriptor/Qualifier:
Animals
Artificial Gene Fusion
Aspergillosis / microbiology,  pathology
Aspergillus fumigatus / drug effects*,  genetics,  growth & development*
Fireflies / genetics
Fungal Proteins / genetics
Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
Luciferases / biosynthesis*,  genetics
Luminescence
Lung / microbiology,  pathology
Male
Mice
Mice, Inbred BALB C
Microbial Sensitivity Tests / methods
Molecular Sequence Data
Promoter Regions, Genetic
Staining and Labeling / methods*
Chemical
Reg. No./Substance:
0/Fungal Proteins; EC 1.13.12.-/Luciferases; EC 1.2.1.-/Glyceraldehyde-3-Phosphate Dehydrogenases
Comments/Corrections

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