Rote N S, Taylor N L, Shigeoka A O, Scott J R, Hill H R
Infect Immun. 1980 Jan;27(1):118-23. doi: 10.1128/iai.27.1.118-123.1980.
We report here on the development of enzyme-linked immunosorbent assays (ELISAs) for antibodies to types II and III group B streptococci. Streptococcal antigens were prepared by trichloroacetic acid extraction and fractional alcohol precipitation. Microtiter wells were coated with antigen in 0.1 M carbonate buffer at pH 9.6. Lyophilization was found to be an essential step for efficient binding of the streptococcal antigens. After incubation with antibody-containing rabbit serum, bound antibody was detected with peroxidase-labeled goat anti-rabbit immunoglobulin G. Optimal antigen concentrations were 200 micrograms/ml for type II and 100 micrograms/ml for type III. An ELISA is also described that uses intact bacteria as antigen. Hyperimmune rabbit serum reacted at a titer in excess of 512 against trichloroacetic acid-soluble antigen and 4,096 against whole bacteria. Sera from human subjects were also tested. Most human sera contained antibody which bound to intact bacteria but not to trichloroacetic acid-solubilized streptococcal antigens. Antibody titers in human sera against intact bacteria correlated very well with opsonic antibody activity measured in a chemiluminescence assay.
我们在此报告针对B族链球菌II型和III型抗体的酶联免疫吸附测定（ELISA）的开发情况。通过三氯乙酸提取和分级乙醇沉淀制备链球菌抗原。微量滴定孔用pH 9.6的0.1 M碳酸盐缓冲液中的抗原包被。发现冻干是链球菌抗原有效结合的关键步骤。与含抗体的兔血清孵育后，用过氧化物酶标记的山羊抗兔免疫球蛋白G检测结合的抗体。II型的最佳抗原浓度为200微克/毫升，III型为100微克/毫升。还描述了一种使用完整细菌作为抗原的ELISA。超免疫兔血清对三氯乙酸可溶性抗原的效价超过512，对全菌的效价为4096。也对人类受试者的血清进行了检测。大多数人血清中含有与完整细菌结合但不与三氯乙酸溶解的链球菌抗原结合的抗体。人血清中针对完整细菌的抗体效价与化学发光测定中测得的调理素抗体活性非常相关。
