首页> 美国卫生研究院文献>Canadian Journal of Comparative Medicine >Immunohistochemical detection of feline calicivirus in formalin-fixed paraffin-embedded specimens.
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Immunohistochemical detection of feline calicivirus in formalin-fixed paraffin-embedded specimens.

机译:在福尔马林固定石蜡包埋的标本中猫杯状病毒的免疫组织化学检测。

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摘要

An immunohistochemical technique was developed for detection of feline calicivirus (FCV) in formalin-fixed, paraffin-embedded cultured cells and tissues. Initial trials with cultured cells indicated that the indirect immunoperoxidase method using rabbit antiserum to FCV strain 255, and horseradish peroxidase-labelled antibodies to rabbit immunoglobulin G lacked sensitivity and showed excessive diffuse background staining despite trypsin digestion of sections before staining. An amplified indirect immunoperoxidase technique using commercially available biotinylated antirabbit antibodies and avidin-biotin-peroxidase or streptavidin-peroxidase (SP) complexes proved highly successful. When optimal conditions, including those for trypsinization, inactivation of endogenous peroxidase and blocking were determined, the SP technique was preferred. Applied to tissue of cats in the acute phase of FCV infection, the technique provided clear identification of cells containing FCV antigens in sections in which histological detail was well preserved.
机译:开发了一种免疫组织化学技术,用于在福尔马林固定,石蜡包埋的培养细胞和组织中检测猫杯状病毒(FCV)。对培养细胞的初步试验表明,使用兔抗FCV 255株抗血清的间接免疫过氧化物酶方法和辣根过氧化物酶标记的兔免疫球蛋白G抗体缺乏敏感性,并且尽管在染色前用胰蛋白酶消化了切片,但仍显示出过度的弥散背景染色。使用市售的生物素化抗兔抗体和抗生物素蛋白-生物素-过氧化物酶或链霉亲和素-过氧化物酶(SP)复合物的扩增间接免疫过氧化物酶技术被证明是非常成功的。如果确定了最佳条件,包括用于胰蛋白酶消化,内源性过氧化物酶的失活和封闭的条件,则首选SP技术。该技术应用于FCV感染急性期的猫组织,可在组织学细节得到很好保存的切片中清楚地鉴定出包含FCV抗原的细胞。

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