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Isolation and Selection of Entomopathogenic Fungi from Soil Samples and Evaluation of Fungal Virulence against Insect Pests
Published on: September 28, 2021
Capillary electrophoresis of conidia from cultivated microscopic filamentous fungi
Marie Horká1, Filip Růzicka, Anna Kubesová
1Institute of Analytical Chemistry Academy of Sciences of the Czech Republic, 602 00 Brno, Czech Republic. horka@iach.cz
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.
Insights
Early detection of fungal infections is crucial for immunocompromised patients. This study introduces a rapid method using capillary electrophoresis to identify fungal conidia, improving diagnostic speed and accuracy.
Area of Science:
- Mycology
- Analytical Chemistry
- Biochemistry
Background:
- Fungal infections pose significant risks, especially in immunocompromised individuals, leading to high mortality rates.
- Current diagnostic methods for fungal pathogens are often time-consuming and labor-intensive.
- Fungi are also implicated in toxic exposures and allergic reactions.
Purpose of the Study:
- To develop and optimize a rapid detection and identification method for filamentous fungi.
- To improve early diagnosis of fungal infections for better patient outcomes.
- To explore advanced separation techniques for fungal conidia analysis.
Main Methods:
- Hydrophobic conidia from filamentous fungi strains were isolated and separated using capillary zone electrophoresis and capillary isoelectric focusing.
- Conidia were dynamically modified with a pyrenebutanoate-based non-ionic tenside for optimized detection.
- Fluorometric detection allowed for the identification of as few as 10 labeled conidia.
Main Results:
- The optimized method enabled the detection of down to 10 labeled fungal conidia.
- Isoelectric points of fungal conidia were successfully determined.
- Separation profiles from clinical samples showed no host specificity, indicating broad applicability.
Conclusions:
- Capillary electrophoresis techniques offer a rapid and sensitive approach for fungal conidia detection.
- This method has the potential to significantly shorten diagnostic timelines for fungal infections.
- The non-host-specific nature of the identified isoelectric points suggests a universal application in fungal diagnostics.
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