Bono J L, Legendre A M, Scalarone G M
Department of Biological Sciences, Idaho State University, Pocatello 83209, USA.
J Med Vet Mycol. 1995 Jul-Aug;33(4):209-14.
Blastomyces dermatitidis (dog isolate T-58) yeast phase lysate antigen was concentrated and separated by Rotofor preparative isoelectric focusing cell (Bio-Rad). The pH values of the fractions were determined and equilibrated to pH 7.2 and then analysed by enzyme-linked immunosorbent assay using horseradish peroxidase enzyme system against serum specimens from dogs with blastomycosis, histoplasmosis, aspergillosis, and coccidioidomycosis. The results showed a peak absorbance at pH 3.89-4.31 (fractions 4 and 5) with the blastomycosis serum specimens. This was a single sharp peak while the rest of the fractions were lower. In contrast the sera from dogs with histoplasmosis showed a peak absorbance at pH 5.54-5.97 (fractions 9 and 10), while the other mycoses showed patterns that did not resemble the blastomycosis or histoplasmosis specimens. Serum specimens from dogs with blastomycosis being treated with itraconazole were also assayed (pre-treatment and 1, 2, 3, and 12 months post-treatment sera). The characteristic peak for blastomycosis was observed and a decrease in the peak was seen as the treatment progressed. Fractions 3-12 were also used to detect delayed dermal hypersensitivity in hyperimmunized hairless guinea-pigs. Fraction 5 (pH 4.31) elicited the optimal response in B. dermatitidis-immunized animals, while no cross-reactivity was observed in guinea-pigs sensitized with Histoplasma capsulatum killed cells.
皮炎芽生菌（犬分离株T - 58）酵母相裂解物抗原通过Rotofor制备型等电聚焦细胞（伯乐公司）进行浓缩和分离。测定各组分的pH值，并将其平衡至pH 7.2，然后使用辣根过氧化物酶酶系统通过酶联免疫吸附测定法对患有芽生菌病、组织胞浆菌病、曲霉病和球孢子菌病的犬血清标本进行分析。结果显示，芽生菌病血清标本在pH 3.89 - 4.31（组分4和5）处出现吸光度峰值。这是一个单一的尖锐峰，而其余组分的吸光度较低。相比之下，患有组织胞浆菌病的犬血清在pH 5.54 - 5.97（组分9和10）处出现吸光度峰值，而其他真菌病呈现出与芽生菌病或组织胞浆菌病标本不同的模式。还对接受伊曲康唑治疗的芽生菌病犬的血清标本进行了检测（治疗前以及治疗后1、2、3和12个月的血清）。观察到了芽生菌病的特征性峰值，并且随着治疗进展峰值降低。组分3 - 12还用于检测超免疫无毛豚鼠的迟发性皮肤超敏反应。组分5（pH 4.31）在经皮炎芽生菌免疫的动物中引发了最佳反应，而在用荚膜组织胞浆菌死细胞致敏的豚鼠中未观察到交叉反应。
