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Identification of Toxoplasma Gondii and Encephalitozoon Cuniculi by Immunoperoxidase Techniques and Electron Microscopy, in Stored, Formalin-Fixed, Paraffin-Embedded Tissue
Identifisering av Toxoplasma gondii og Encephalitozoon cuniculi ved hjelp av immunoperoksydase-teknikk og elektronmikroskopi i lagret, formalin-fiksert, parafin-innstøpt vevsmateriale
Acta Veterinaria Scandinavica volume 27, pages 11–22 (1986)
Abstract
The peroxidase-anti-peroxidase (PAP), avidin-biotin-complex (ABC) techniques, and electron microscopy, were used to identify protozoan parasites in formalin-fixed material from routine necropsy cases. The material comprised paraffin blocks, that had been stored for up to 18 years, from 18 cases of suspected toxoplasmosis and encephalitozoonosis. With the ABC method a higher dilution of primary antibody could be used than with the PAP method. However, with both methods, a distinct reaction occurred with appropriate dilutions. The age of the material did not seem to effect the result. Frequently, phagocytized and necrotic parasites were also stained. Gystozoites (bradyzoites) of T. gondii were stained faintly compared to the endozoites (tachyzoites). A Toxoplasma-like parasite from a dog did not react with anti-Toxoplasma serum, and ultrastructurally it proved to be consistent with an unidentified cyst-forming sporozoan parasite previously reported in dogs. Electron microscopy based on paraffin-embedded tissue, seems to be a valuable method for identification of protozoan parasites, and thus provide a supplement or alternative to the immunoperoxidase methods.
Sammendrag
Peroksydase-antiperoksydase- (PAP) og avidin-biotin-kompleks-teknikk (ABG) ble benyttet til å identifisere protozoer i formalinfiksert materiale. Materialet besto av parafinblokker fra 18 kasus hvor det var mistanke om toxoplasmose eller encephalitozoonose. Enkelte blokker hadde vært lagret så lenge som 18 år. ABG-metoden var mer sensitiv enn PAP-metoden, men ved passende fortynning av primært antistoff ble det tydelig reaksjon ved begge metodene. Materialets alder syntes ikke å påvirke resultatet. Fagocytterte og nekrotiske parasitter ble også ofte farget. Cystozoitter (bradyzoitter) av T. gondii ble farget svakere enn endozoitter (tachyzoitter). En Toxoplasma-lignende parasitt fra hund reagerte ikke med anti-Toxoplasma serum. Elektronmikroskopisk undersøkelse tydet imidlertid på at den var identisk med en cyste-dannende sporozo som er beskrevet tidligere hos hund. Elektronmikroskopi basert på parafin-innstøpt materiale ser ut til å være en egnet metode for identifisering av protozoer og representerer således et supplement eller alternativ til immunoperoksydase-teknikkene.
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Bjerkås, I., Landsverk, T. Identification of Toxoplasma Gondii and Encephalitozoon Cuniculi by Immunoperoxidase Techniques and Electron Microscopy, in Stored, Formalin-Fixed, Paraffin-Embedded Tissue. Acta Vet Scand 27, 11–22 (1986). https://doi.org/10.1186/BF03548555
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DOI: https://doi.org/10.1186/BF03548555