Shimizu S, Shimura K, Ito S, Minami T
Laboratory of Protozoa, National Institute of Animal Health, Tsukuba Science City, Japan.
Parasitol Res. 1992;78(8):684-8. doi: 10.1007/BF00931521.
A method of isolating Babesia ovata merozoites from infected erythrocytes and an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-B. ovata antibodies were developed. After B. ovata-infected erythrocytes had been lysed using the nitrogen cavitation method, the merozoites were separated from erythrocyte components by differential centrifugation and density-gradient centrifugation. The light microscopic examination showed that the purified merozoites were morphologically intact and contained few contaminants. Sodium dodecyl sulfate-polyacrylamide electrophoretic (SDS-PAGE) analysis revealed that the merozoite fraction contained very little contamination with erythrocyte components. The merozoites thus obtained were sonicated and treated with Triton X-100 and then used as an antigen to measure anti-B. ovata antibodies in ELISA. The ELISA was more sensitive in detecting anti-B. ovata antibodies than was either the indirect fluorescent antibody test or the complement fixation test on sera from cattle infected with B. ovata.
开发了一种从感染红细胞中分离卵形巴贝斯虫裂殖子的方法以及一种用于检测抗卵形巴贝斯虫抗体的酶联免疫吸附测定（ELISA）。使用氮空化法裂解卵形巴贝斯虫感染的红细胞后，通过差速离心和密度梯度离心从红细胞成分中分离出裂殖子。光学显微镜检查表明，纯化的裂殖子形态完整且几乎没有污染物。十二烷基硫酸钠-聚丙烯酰胺电泳（SDS-PAGE）分析显示，裂殖子部分几乎没有被红细胞成分污染。将如此获得的裂殖子进行超声处理并用Triton X-100处理，然后用作抗原在ELISA中测量抗卵形巴贝斯虫抗体。ELISA在检测抗卵形巴贝斯虫抗体方面比间接荧光抗体试验或对感染卵形巴贝斯虫的牛血清进行的补体结合试验更敏感。
