Improved detection of medically important fungi by immunoperoxidase staining with polyclonal antibodies

Abstract

This study was performed to identify pathological fungi of eight species [Aspergillus fumigatus, Candida albicans, Torulopsis (Candida) glabrata, Cryptococcus neoformans, Fusarium anthophilum, Rhizopus oryzae, Sporothrix schenckii and Trichosporon beigelii] in formalin-fixed, paraffin-embedded tissue sections by indirect immunoperoxidase staining. Mature albino rabbits were immunized with formalin-killed organisms. Antibodies were prepared by precipitation. Immunoperoxidase staining was applied to the paraffin-embedded tissue sections of experimentally infected mice and human autopsy and surgical specimens. Although the cell walls of each fungus stained clearly, many cross-reactivities appeared. However, it was possible to obtain specificity for the eight species by absorption and dilution of the antisera.

Utilização de três métodos imuno-histoquímicos na detecção de aspergilose e zigomicose em animais

“…A ausência de imunomarcação nos conidióforos, observada nos casos de aspergilose, pode estar associada à variação na capacidade de imunomarcação dos elementos fúngicos devido às diferenças existentes na composição antigênica nas diferentes fases de desenvolvimentos dos fungos (Fukuzawa et al 1995).…”

“…nos casos de aspergilose, zigomicose, candidíase e pitiose aqui testados, demonstrando uma alta especificidade desses anticorpos. Entretanto, diversos estudos realizados com determinados anticorpos anti-antígenos fungo-específicos têm demonstrado reações cruzadas na IHQ (Reed et al 1993, Fukuzawa et al 1995, Verweij et al 1996, Schuetz & Cohen 2009). Nesses casos, as reações cruzadas podem estar associadas às semelhanças antigênicas entre alguns fungos (Reed et al 1993, Fukuzawa et al 1995, Schuetz & Cohen 2009).…”

“…Entretanto, diversos estudos realizados com determinados anticorpos anti-antígenos fungo-específicos têm demonstrado reações cruzadas na IHQ (Reed et al 1993, Fukuzawa et al 1995, Verweij et al 1996, Schuetz & Cohen 2009). Nesses casos, as reações cruzadas podem estar associadas às semelhanças antigênicas entre alguns fungos (Reed et al 1993, Fukuzawa et al 1995, Schuetz & Cohen 2009). Na medida em que novos anticorpos (livres de reações cruzadas) são desenvolvidos e testados através da IHQ (Marcilla et al 1999), esta técnica vai se apresentando como uma alternativa de diagnóstico específica e eficiente diante de testes mais onerosos e que não combinam a morfologia com a detecção do fungo específico (Guarner & Brandt 2011).…”

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“…There have also been some successful attempts at detecting circulating antibodies (IgG) from infected patients by serological assays such as ELISA and the precipitin-in-gel method [4,5]. Various staining methods such as immunoperoxidase staining [6] and Gomori's methenamine silver (GMS) stain [2] are also used for detecting pathogens. The use of other diagnostic methods such as auto¯uorescence [7], PCR [8], thin-layer chromatography and high-performance liquid chromatography for toxin detection [9], SDS-PAGE and Western blotting procedures [10] has been reported.…”

Development of a Peptide Nucleic Acid Probe to Trichosporon Species and Identification of Trichosporonosis by Use of In Situ Hybridization in Formalin-Fixed and Paraffin-Embedded (FFPE) Sections

“…The diagnosis of trichosporonosis by immunohistochemistry using a self-made antibody to Trichosporon has been reported (18,19); however, these antibodies are not available for commercial use, there are limitations for their use, and their specificity could not be confirmed. Therefore, we developed the auxiliary utility of ISH for the pathological diagnosis of trichosporonosis from FFPE tissues.…”