Capillary Electrophoresis of Conidia from Cultivated Microscopic Filamentous Fungi

Abstract

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.

Preparative and capillary isoelectric focusing for detection and identification of Aspergillus conidia in complex sample matrices

“…The electropherograms are shown in Figure . The found pI values of conidia reported in were confirmed, and were determined as 2.05 for A. niger , 3.35 for A. parasiticus , 5.90 for A. flavus (Figure A), and 3.15 for A. fumigatus (Figure B). The sample of A. fumigatus conidia was injected alone into the capillary for greater clarity of the analysis in the selected wide pH gradient.…”

“…In , the conidia of filamentous fungi (one type of the conidia), A. niger , A. fumigatus , A. flavus , Fusarium solani , and Penicillium chrysogenum were separated according to their pI and electrophoretic mobilities by CIEF and CZE, respectively. The use of capillary techniques is limited by tens of nanoliters of a homogeneous sample which can be injected directly into the separation capillary.…”

“…Fungi can produce carcinogenic mycotoxins that are important etiological agents of nosocomial infections in immunocompromised patients. Morphological and physiological characteristics of fungal conidia can be used to identify the original strains of fungi . Pathogenic conidia can elicit an allergic response resulting in the development of rhinitis or asthma by their inhalation.…”

Latest improvements in CIEF: From proteins to microorganisms

“…In recent years, several studies focused on CIEF separations of different filamentous fungi were published. Conidia from cultures of different strains of filamentous fungi together with several strains isolated from clinical material, native or labeled by PB‐PEG, were analyzed by CIEF with UV and fluorometric detection . In accordance with previous studies dealing with bacteria or yeasts, determined p I values were found independent of the host origin and were not affected by the modification with PB‐PEG.…”

Highly accurate deterministic lateral displacement device and its application to purification of fungal spores

“…7 An inertial microfluidic device was recently shown to separate E. coli from red blood cells 8 and immunomagnetic separation can be used for smaller numbers of organisms. 9 Capillary electrophoresis was recently used to separate conidia ͑spores͒ from various filamentous fungi including Aspergillus, 10 but there are few microfluidic systems that demonstrated application to fungal biology.…”