Palmer G H, Barbet A F, Kuttler K L, McGuire T C
J Clin Microbiol. 1986 Jun;23(6):1078-83. doi: 10.1128/jcm.23.6.1078-1083.1986.
The establishment and maintenance of anaplasmosis-free cattle herds is impaired due to the lack of a rapid, sensitive, and specific serologic test to detect persistently infected cattle which serve as carriers for the organism. To develop an improved diagnostic test for anaplasmosis we screened Anaplasma marginale initial body proteins to identify a protein common to antigenically different isolates that is recognized by the host immune system at all stages of infection. Seronegative cattle were infected with either the Florida, Virginia, or North Texas isolate of A. marginale and monitored for infection by daily examination of Wright-stained blood smears for parasitized erythrocytes. Sera from cattle at different stages of infection, from acute through persistent, were used to immunoprecipitate A. marginale proteins that were metabolically radiolabeled with [35S]methionine or surface radiolabeled with 125I. Multiple A. marginale proteins were recognized by using sera either undiluted or at 1:10; however, only four or five proteins were sufficiently antigenic to elicit antibody reactive with a 1:1,000 serum dilution. A single protein with an apparent molecular mass of 86 kilodaltons was consistently recognized at all stages of infection regardless of the isolate used to infect the cattle. This protein was demonstrated to be on the surface of the A. marginale initial body and to be water soluble. We propose use of this 86-kilodalton protein to develop an improved serologic test for diagnosis of bovine anaplasmosis.
由于缺乏一种快速、灵敏且特异的血清学检测方法来检测持续感染的牛（这些牛是该病原体的携带者），无嗜吞噬细胞无形体病牛群的建立和维持受到了阻碍。为了开发一种改进的嗜吞噬细胞无形体病诊断检测方法，我们筛选了边缘无浆体初始体蛋白，以鉴定一种在抗原性不同的分离株中都存在的蛋白，该蛋白在感染的各个阶段都能被宿主免疫系统识别。血清学阴性的牛感染了边缘无浆体的佛罗里达、弗吉尼亚或北德克萨斯分离株，并通过每天检查经瑞氏染色的血涂片以寻找被寄生的红细胞来监测感染情况。来自处于从急性感染到持续感染不同阶段的牛的血清，用于免疫沉淀经[35S]甲硫氨酸代谢性放射性标记或经125I表面放射性标记的边缘无浆体蛋白。使用未稀释或1:10稀释的血清可识别多种边缘无浆体蛋白；然而，只有四五种蛋白具有足够的抗原性，能引发与1:1000血清稀释度反应的抗体。无论用于感染牛的分离株如何，一种表观分子量为86千道尔顿的单一蛋白在感染的各个阶段都能持续被识别。该蛋白被证明位于边缘无浆体初始体的表面且可溶于水。我们建议使用这种86千道尔顿的蛋白来开发一种改进的血清学检测方法，用于诊断牛嗜吞噬细胞无形体病。
