Detection of Aspergillus fumigatus hyphae by solid phase cytometry

Vos, M.M. De; Nelis, Hans

doi:10.1016/s0167-7012(03)00174-x

Cited by 14 publications

(4 citation statements)

References 16 publications

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“…and they may be easily influenced by subjective error in results evaluation. Currently, some new methods − appeared, for example, the rapid identification and characterization products and antigens of FF − and methods based on detection of nucleic acids by polymerase chain reaction (PCR) . The reliability of the methods is high.…”

mentioning

confidence: 99%

Capillary Electrophoresis of Conidia from Cultivated Microscopic Filamentous Fungi

et al. 2009

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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.

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confidence: 99%

Capillary Electrophoresis of Conidia from Cultivated Microscopic Filamentous Fungi

et al. 2009

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“…Previously, some studies have used SPC and/or immunostaining for the detection of A. fumigatus. However, these studies either missed the quantitative aspect, as they used standard microscopy for detection (37), or depended on the validation step to detect the fluorescence obtained by immunostaining (38,39). In the present study, the combination of immunodetection and the TSA method improved the fluorescence intensity, enabling direct detection of the labeled A. fumigatus microcolonies by the solid-phase cytometer.…”

Section: Discussionmentioning

confidence: 76%

Rapid Detection and Quantification of Aspergillus fumigatus in Environmental Air Samples Using Solid-Phase Cytometry

Vanhee

Nelis

Coenye

2009

Environ. Sci. Technol.

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Aspergillus fumigatus is an ubiquitous fungus capable of causing severe infections such as aspergilloma, allergic bronchopulmonary aspergillosis, and invasive aspergillosis, especially in immunocompromised patients. Monitoring the number of Aspergillus fumigatus spores in the air is crucial for infection control. In the present study, a novel approach for the quantification of Aspergillus fumigatus, based on solid-phase cytometry (SPC) and immunofluorescent labeling, was developed. The sensitivity and specificity of the assay were confirmed by testing pure cultures. Paecilomyces variotii and Rhizopus stolonifer were codetected but could be excluded on the basis of morphology of the microcolonies. The SPC method has considerable advantages compared to the culture-based method, including its low detection limit (4 cells/m3), its speed (results are obtained within 24 h), and the straightforward microscopic identification of Aspergillus fumigatus. Additionally, comparison of results obtained with both methods demonstrated that they are equally accurate for the quantification of Aspergillus fumigatus in environmental air samples.

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“…• Specific detection of A. fumigatus in bronchoalveolar lavage liquid and sputum by combining viability staining and immunofluorescence labelling 20,21 .…”

Section: Applications Of Spcmentioning

confidence: 99%

Detection and quantification of viable airborne bacteria and fungi using solid-phase cytometry

2009

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This protocol describes the use of solid-phase cytometry for the enumeration of airborne bacteria and fungi. In contrast with conventional methods, accurate results can be obtained in real time, especially for air samples with low numbers of microorganisms. Air samples are collected by impaction on a water-soluble polymer that is subsequently dissolved. Part of the sample can be filtered over two membrane filters with different pore sizes. One filter is used to obtain a total count of all viable microorganisms, and a second filter is used to determine the number of airborne fungi. Microorganisms present on the filter are labeled with a viability substrate and subsequently detected and quantified using a solid-phase cytometer. The detected spots are microscopically validated using an epifluorescence microscope to discriminate between bacteria, fungi and fluorescent particles. The whole procedure takes 5 h to complete and results in the accurate quantification of airborne bacteria and fungi for samples with a low or high microbial load

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Detection of Aspergillus fumigatus hyphae by solid phase cytometry

Cited by 14 publications

References 16 publications

Capillary Electrophoresis of Conidia from Cultivated Microscopic Filamentous Fungi

Capillary Electrophoresis of Conidia from Cultivated Microscopic Filamentous Fungi

Rapid Detection and Quantification of Aspergillus fumigatus in Environmental Air Samples Using Solid-Phase Cytometry

Detection and quantification of viable airborne bacteria and fungi using solid-phase cytometry

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