Optimizing immunohistochemical staining for rapid diagnosis of porcine epidemic diarrhea

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  • Yaguchi Y.
    Ibaraki Prefectural Kenpoku Livestock Hygiene Service Center
  • Takahashi S.
    Ibaraki Prefectural Kensei Livestock Hygiene Service Center
  • Komatsu Y.
    Ibaraki Prefectural Kennan Livestock Hygiene Service Center
  • Kawashima K.
    National Institute of Animal Health, National Agricultural and Food Research Organization

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  • 豚流行性下痢の迅速診断に適した免疫組織化学的染色条件の検討
  • ブタ リュウコウセイ ゲリ ノ ジンソク シンダン ニ テキシタ メンエキ ソシキ カガクテキ センショク ジョウケン ノ ケントウ

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Abstract

<p>Porcine epidemic diarrhea（PED）is an acute viral disease characterized by watery diarrhea and high mortality in newborn piglets. Immunohistochemical（IHC）detection of PED virus on the intestinal tissue is effective for the diagnosis of PED. However, IHC diagnosis of PED using routine fixation is time-consuming. We therefore investigated the usefulness of a rapid fixation method and frozen sections using a field sample for faster diagnosis of PED. For rapid fixation, the tissues were fixed in 10% neutral buffered formalin for different time periods and at different temperatures and compared for staining pattern and intensity. For frozen sections, tissues were frozen in hexane at -80℃, sectioned and fixed in formalin-methanol. The sections were then processed by microwave heating for antigen retrieval and immunostained for PED antigens. As a result, the rapid fixation at 40 and 60℃ for 1 h was sufficient for the detection of PED virus. By this rapid fixation method, the results of IHC diagnosis can be obtained by the following day. The cryosections also exhibited specific immunostaining and well-preserved morphology. In conclusion, these rapid fixation and frozen section protocols have a potential use for the rapid diagnosis of PED within a day or two of necropsy.</p>

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