泰氏锥虫抗原制备及实验室诊断研究

Studies on the Preparation of Antigen of Trypanosoma theileri and Apply them to Laboratory Diagnosis

应用体外培养的泰氏锥虫制备可溶性抗原Ⅰ、抗原Ⅱ和代谢抗原.经测定,其蛋白含量每毫升分别为6.5mg、7.4mg和7.1mg.薄层等电聚焦电泳测定结果表明,抗原Ⅰ出现22条区带,抗原Ⅱ21条区带,代谢抗原28条区带,对照的伊氏锥虫琼脂免疫扩散抗原14条区带.经分析,泰氏锥虫抗原和伊氏锥虫抗原有4条区带在同一迁移率上.琼脂免疫扩散反应中,3种泰氏锥虫抗原均与相应免疫兔血清发生沉淀反应,抗原Ⅰ出现1条致密沉淀线,抗原Ⅱ和代谢抗原出现2～3条沉淀线,抗原效价为1:4～16.免疫电泳显示了类似的结果,抗原Ⅰ与免疫兔血清出现1条弧形沉淀线,抗原Ⅱ和代谢抗原与免疫兔血清出现了3条弧形沉淀线.间接血凝试验结果表明,泰氏锥虫自然感染牛血清效价为1:20～40,免疫兔血清为1:1280～5120.所制泰氏锥虫抗原对伊氏锥虫和媾疫锥虫血清也能很好地发生交叉反应,3份伊氏锥虫病马血清和3份伊氏锥虫人工感染兔血清血凝效价分别为1:10～40和1:8～1024;5份媾疫马血清有4份血凝效价为1:20～320.4份环形泰勒虫病牛血清均为阴性.

Soluble antigen F, 222222 and metabolic antigen were prepared from cultures of Try-panosoma theiieri in vitro,which having 6. 5 mg/mL,7. 4 mg/mL and 7. 1mg/mL of proteins respectively. The results from isoelectric focusing showed that the antigen 11111I , 222222 and metabolic antigen revealed 22,21 and 28 zones respectively, while T. evansi antigen used as control gave rise to 14 zones in agar gel diffusion. There were 4 zones having identical migration rates among antigens of T. theiieri and T. evansi. In agar gel diffusion, all these antigens reacted with the corresponding immune sera from rabbits,and antigen I formed 1 dense precipitating line and the remaining two antigens produced 2-3 lines. The titers of these antigens were 1 : 4 to 1 : 16. The similar results were obtained with immunoelectrophoresis. Indirect haemagglutination showed that the titers of sera from cattle naturally infected with T. theiieri was 1 : 20 to 1 : 40 and those of immunized rabbits' sera was 1 : 1280 to 1 : 2560. The prepared antigens could efficiently cross-react with sera of animals infected with T. evansi or T. equiperdum. The indirect haemagglutinating titers of 3 serum samples from horses with surra and 3 serum samples of rabbits experimentally infected with T. evansi were 1 : 10 to 1 : 40 and 1 : 8 to 1 : 1024,respectively;4 out of 5 serum samples of horses with dourine exhibited 1 : 20 to 1 : 320 of indirect haemagglutinating titers. Four serum samples of cattle infected with Theileria annulata failed to show positive reaction.