Montenegro-James S, Guillen A T, Ma S J, Tapang P, Abdel-Gawad A, Toro M, Ristic M
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Illinois, Urbana 61801.
Am J Vet Res. 1990 Oct;51(10):1518-21.
Isolated Anaplasma marginale initial bodies were successfully used in a dot ELISA for rapid detection of antibodies to Anaplasma organisms. The enzyme immunoassay used only 25 ng of antigen dotted onto nitrocellulose disks. Antigen-antibody complexes were detected by use of alkaline phosphatase-conjugated protein A, and reactions were read visually after addition of a precipitable, chromogenic substrate. The test allowed the processing of multiple sera, either for screening or for titer determination, in less than 3 hours and was found to be as sensitive as the indirect fluorescent antibody test. The overall performance of the dot ELISA, using isolated A marginale initial bodies, for 580 bovine serum samples was as follows: sensitivity, 93%; specificity, 96%; and predictive value, 95%. Cross-reactivity was not observed with sera positive to Babesia bovis and B bigemina, Trypanosoma vivax, or common bacteria or viruses infecting cattle. The antigen dotted onto nitrocellulose disks was stable when stored at -20, 4, or 25 C. Compared with the indirect fluorescent antibody test, the dot ELISA allowed easier, faster, and more objective interpretation of results. Its simplicity and low cost combined with high sensitivity and specificity indicate that this assay could effectively replace serologic assays currently used for diagnosis of anaplasmosis in cattle.
将分离出的边缘无形体原始小体成功用于斑点酶联免疫吸附测定（dot ELISA），以快速检测针对无形体生物体的抗体。该酶免疫测定仅使用点样于硝酸纤维素圆盘上的25 ng抗原。抗原 - 抗体复合物通过使用碱性磷酸酶偶联的蛋白A进行检测，在加入可沉淀的显色底物后目视读取反应结果。该检测方法能够在不到3小时内处理多个血清样本用于筛查或滴度测定，并且发现其与间接荧光抗体试验一样灵敏。使用分离出的边缘无形体原始小体进行的斑点酶联免疫吸附测定对580份牛血清样本的总体性能如下：敏感性为93%；特异性为96%；预测值为95%。未观察到与牛巴贝斯虫、双芽巴贝斯虫、活泼锥虫或感染牛的常见细菌或病毒呈阳性反应的血清发生交叉反应。点样于硝酸纤维素圆盘上的抗原在-20℃、4℃或25℃储存时稳定。与间接荧光抗体试验相比，斑点酶联免疫吸附测定能够更简便、快速且客观地解读结果。其简单性和低成本，再加上高敏感性和特异性，表明该检测方法可有效替代目前用于牛无浆体病诊断的血清学检测方法。
