McCorkle-Shirley S, Hart L T, Larson A D, Todd W J, Myhand J D
Am J Vet Res. 1985 Aug;46(8):1745-7.
Purification of Anaplasma marginale from infected bovine RBC was achieved through enzyme treatment and density-gradient centrifugation. A relative yield of 41.6% was obtained by dividing the number of organisms in the final purified preparation by the number of A marginale-infected RBC. Purified parasites were verified as A marginale by light microscopy, electron microscopy, and immunologic tests. The purified parasites reacted positively with calf and rabbit anti-A marginale sera in interfacial and slide agglutination tests. Anti-bovine RBC serum did not agglutinate purified A marginale, indicating absence of any contaminating RBC stroma. Anaplasma marginale was antigenic, but did not cause infection when the preparation was inoculated into a susceptible calf. The density of A marginale was determined to be 1.19 g/ml and cell diameters ranged from 0.25 to 0.63 micron. This method provided procedures for obtaining A marginale free of bovine RBC antigens for accurate biochemical assays and vaccine production.
通过酶处理和密度梯度离心法从感染牛红细胞中纯化出边缘无形体。通过将最终纯化制剂中的微生物数量除以边缘无形体感染的红细胞数量，获得了41.6%的相对产量。通过光学显微镜、电子显微镜和免疫测试，验证纯化的寄生虫为边缘无形体。纯化的寄生虫在界面和玻片凝集试验中与小牛和兔抗边缘无形体血清呈阳性反应。抗牛红细胞血清不会凝集纯化的边缘无形体，表明不存在任何污染的红细胞基质。边缘无形体具有抗原性，但当将制剂接种到易感小牛体内时不会引起感染。边缘无形体的密度测定为1.19 g/ml，细胞直径范围为0.25至0.63微米。该方法提供了获得不含牛红细胞抗原的边缘无形体的程序，用于准确的生化分析和疫苗生产。
