Specific detection of fusarium species in blood and tissues by a PCR technique.
Fusarium species are opportunistic nosocomial pathogens that often cause fatal invasive mycoses. We designed a primer pair that amplifies by PCR a fragment of a gene coding for the rRNA of Fusarium species. The DNAs of the main Fusarium species and Neocosmospora vasinfecta but not the DNAs from 11 medically important fungi were amplified by these primers. The lower limit of detection of the PCR system was 10 fg of Fusarium solani DNA by ethidium bromide staining. To test the ability of this PCR system to detect Fusarium DNA in tissues, we developed a mouse model of disseminated fusariosis. Using the PCR, we detected Fusarium DNA in mouse tissues and in spiked human blood. Furthermore, F. solani, Fusarium moniliforme, and Fusarium oxysporum were testing by random amplified polymorphic DNA (RAPD) analysis. The bands produced by RAPD analysis were purified, cloned, and sequenced. The information was used to design primer pairs that selectively amplified one or several Fusarium species. The method developed may be useful for the rapid detection and identification of Fusarium species both from culture and from clinical samples.
Laboratoire de Mycologie Médicale, Institut Pasteur, 75724 Paris cedex 15, France. cneyret@ipagua.gp
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D000818:Animals; D004271:DNA, Fungal / Q000032:analysis* / Q000235:genetics; D005670:Fusarium / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D051379:Mice; D009181:Mycoses / Q000097:blood / Q000382:microbiology*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*
D004271:DNA, Fungal; D015345:Oligonucleotide Probes