Capillary electrophoresis of conidia from cultivated microscopic filamentous fungi.
In immunocompromised people fungal agents are able to cause serious infections with high mortality rate. An early diagnosis can increase the chances of survival of the affected patients. Simultaneously, the fungi produce toxins and they are frequent cause of allergy. Currently, various methods are used for detection and identification of these pathogens. They use microscopic examination and growth characteristic of the fungi. New methods are based on the analysis of structural elements of the target microorganisms such as proteins, polysaccharides, glycoproteins, nucleic acids, etc. for the construction of antibodies, probes, and primers for detection. The above-mentioned methods are time-consuming and elaborate. Here hydrophobic conidia from the cultures of different strains of the filamentous fungi were focused and separated by capillary zone electrophoresis and capillary isoelectric focusing. The detection was optimized by dynamic modifying of conidia by the nonionogenic tenside on the basis of pyrenebutanoate. Down to 10 labeled conidia of the fungal strains were fluorometrically detected, and isoelectric points of conidia were determined. The observed isoelectric points were compared with those obtained from the separation of the cultured clinical samples, and they were found to be not host-specific.
Horká
Marie
M
Institute of Analytical Chemistry Academy of Sciences of the Czech Republic, 602 00 Brno, Czech Republic. horka@iach.cz
Růzicka
Filip
F
Kubesová
Anna
A
Holá
Veronika
V
Slais
Karel
K
D019075:Electrophoresis, Capillary / Q000379:methods*; D005658:Fungi / Q000737:chemistry; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet; D013172:Spores, Fungal / Q000737:chemistry* / Q000302:isolation & purification