Machado R Z, Montassier H J, Pinto A A, Lemos E G, Machado M R, Valadão I F, Barci L G, Malheiros E B
Faculdade de Ciências Agrárias e Veterinárias-UNESP, Rod. Carlos Tonanni, Jaboticabal, São Paulo, Brazil.
Vet Parasitol. 1997 Jul 15;71(1):17-26. doi: 10.1016/s0304-4017(97)00003-4.
A method for the isolation of Babesia bovis merozoites from infected erythrocytes (Machado et al., 1994) and an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-B. bovis antibodies were developed. This ELISA utilizes a soluble, alkali-digested B. bovis antigen. Sera from calves experimentally infected with B. bovis were screened by this technique from day 9 to day 233 postinfection (PI). Maximum titers were reached between days 29 and 149 PI. Sera from calves (n = 62), heifers (n = 38) and cows (n = 49), raised in tick-infested areas of São Paulo State, showed higher antibody levels in heifers and cows. A higher percentage of negative sera (19.4%) was found among calves. Sodium dodecyl sulphate-polyacrylamide electrophoresis (SDS-PAGE) and immunoblotting have identified proteins of similar molecular mass in the two species. Sera from calves experimentally infected with B. bovis reacted with homologous antigens at the level of 95, 66 and 23 kDa. The same serum reacted with the 23 kDa band of heterologous antigen. Sera from calves experimentally infected with B. bigemina recognized 82, 66, 58, 36 and the 23 kDa polypeptides of homologous and heterologous antigens. The experimental ELISA described may prove to be a practical serological test for bovine Babesia infection with the choice of specific test antigen for B. bovis and B. bigemina.
已开发出一种从受感染红细胞中分离牛巴贝斯虫裂殖子的方法（马查多等人，1994年）以及一种用于检测抗牛巴贝斯虫抗体的酶联免疫吸附测定（ELISA）。该ELISA使用可溶性碱消化的牛巴贝斯虫抗原。用该技术对实验感染牛巴贝斯虫的犊牛血清在感染后第9天至第233天进行筛查。在感染后第29天至第149天达到最高滴度。在圣保罗州蜱虫滋生地区饲养的犊牛（n = 62）、小母牛（n = 38）和母牛（n = 49）的血清显示，小母牛和母牛的抗体水平较高。在犊牛中发现较高比例的阴性血清（19.4%）。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹已鉴定出这两个物种中分子量相似的蛋白质。实验感染牛巴贝斯虫的犊牛血清与95、66和23 kDa水平的同源抗原发生反应。同一血清与异源抗原的23 kDa条带发生反应。实验感染双芽巴贝斯虫的犊牛血清识别同源和异源抗原的82、66、58、36和23 kDa多肽。所描述的实验ELISA可能被证明是一种用于牛巴贝斯虫感染的实用血清学检测方法，可选择针对牛巴贝斯虫和双芽巴贝斯虫的特异性检测抗原。
