Jensen H E, Aalbaek B, Lind P, Frandsen P L, Krogh H V, Stynen D
Department of Pharmacology and Pathobiology, Royal Veterinary and Agricultural University, Frederiksberg, Denmark.
APMIS. 1993 Jul;101(7):505-16. doi: 10.1111/j.1699-0463.1993.tb00140.x.
To improve the immunohistopathological diagnosis of systemic bovine mycoses, we have evaluated the utility of antifungal polyclonal and monoclonal antibodies, and peroxidase and alkaline phosphatase staining techniques. A rabbit polyclonal antibody to mannan from Candida albicans was specific for candidosis. The diagnosis of aspergillosis was accomplished using a rat monoclonal antibody to the galactofuran side chains of Aspergillus galactomannan. A murine monoclonal antibody reacting with weakly Con-A binding 41 and 46 kDa somatic antigens from Absidia corymbifera was used for immunostaining of zygomycetic hyphae. Peroxidase antiperoxidase (PAP) and alkaline phosphatase antialkaline phosphatase (APAAP) complexes were visualized using aminoethylcarbazole and fast red substrates. A green staining of PAP reactions with dioctyl sulfosuccinate sodium and 3,3',5,5'-tetramethylbenzidine (DONS/TMB) was effective for the demonstration of fungi in dual and triple infections. Tissue sections of experimentally infected mice were used to determine the sensitivity and specificity of the antibodies. Tissues obtained from 161 bovine mycotic lesions previously studied by indirect immunofluorescence staining were further evaluated using the three antibodies. In all of 45 lesions solely affected by aspergillosis and in three solely affected by candidosis the diagnoses were confirmed by the new evaluation. In 85 of 96 cases of single infections with zygomycetes the diagnosis was confirmed, while none of the antibodies reacted with fungal elements in the remaining 11 lesions. Aspergillus hyphae were detected in all three lesions with dual aspergillosis and zygomycosis, whereas zygomycetic material was confirmed in only two of these cases. A mixed infection of candidosis and zygomycosis in a lymph node was confirmed too. In 13 cases in which a diagnosis had not hitherto been obtained, aspergillosis and zygomycosis were recorded each in three cases.
为提高牛全身性真菌病的免疫组织病理学诊断水平，我们评估了抗真菌多克隆抗体和单克隆抗体以及过氧化物酶和碱性磷酸酶染色技术的效用。一种针对白色念珠菌甘露聚糖的兔多克隆抗体对念珠菌病具有特异性。使用针对烟曲霉半乳甘露聚糖半乳呋喃侧链的大鼠单克隆抗体完成曲霉病的诊断。一种与来自伞枝犁头霉的弱刀豆球蛋白A结合的41和46 kDa体细胞抗原发生反应的小鼠单克隆抗体用于接合菌菌丝的免疫染色。使用氨基乙基咔唑和固红底物使过氧化物酶抗过氧化物酶（PAP）和碱性磷酸酶抗碱性磷酸酶（APAAP）复合物显色。用磺基琥珀酸二辛酯钠和3,3',5,5'-四甲基联苯胺（DONS/TMB）对PAP反应进行绿色染色可有效显示双重和三重感染中的真菌。使用实验感染小鼠的组织切片来确定抗体的敏感性和特异性。使用这三种抗体进一步评估了从先前通过间接免疫荧光染色研究的161例牛真菌性病变中获得的组织。在仅受曲霉病影响的45个病变以及仅受念珠菌病影响的3个病变中，新的评估均证实了诊断。在96例接合菌单感染病例中的85例中诊断得到证实，而其余11个病变中的真菌成分均未与任何抗体发生反应。在所有3例曲霉病和接合菌病双重感染的病变中均检测到曲霉菌丝，而在这些病例中仅2例证实有接合菌物质。还证实了淋巴结中念珠菌病和接合菌病的混合感染。在13例迄今未获得诊断的病例中，曲霉病和接合菌病各记录了3例。
