CAJ | 학술논문

以真核表达的焦虫表面抗原重组蛋白 TaSP-Tams1-SPAG1作为 ELISA 板包被抗原,通过方阵实验确定抗原包被浓度,探索抗原包被时间、温度、一抗及二抗血清稀释度及作用时间,建立牛环形泰勒焦虫表面抗原重组蛋白 ELISA 方法.结果得出 TaSP-Tams1-SPAG1的最佳包被浓度为10μg/mL,血清最佳稀释度为1∶100, ELISA 阳性反应的临界值为 OD 492nm ≥0．348,重复试验变异系数小于12％,所建立的 ELISA 方法不与牛巴贝斯虫及牛瑟氏泰勒虫血清发生反应.TaSP-Tams1-SPAG1为抗原建立的 ELISA 方法特异性强,敏感度高,重复性好,为牛环形泰勒焦虫病的准确检测及防治提供技术支持.
이진핵표체적초충표면항원중조단백 TaSP-Tams1-SPAG1작위 ELISA 판포피항원,통과방진실험학정항원포피농도,탐색항원포피시간、온도、일항급이항혈청희석도급작용시간,건립우배형태륵초충표면항원중조단백 ELISA 방법.결과득출 TaSP-Tams1-SPAG1적최가포피농도위10μg/mL,혈청최가희석도위1∶100, ELISA 양성반응적림계치위 OD 492nm ≥0．348,중복시험변이계수소우12％,소건립적 ELISA 방법불여우파패사충급우슬씨태륵충혈청발생반응.TaSP-Tams1-SPAG1위항원건립적 ELISA 방법특이성강,민감도고,중복성호,위우배형태륵초충병적준학검측급방치제공기술지지.
The objective of this paper was to discuss the antigen coating time,temperature,serum dilution of first antibody and second antibody and action time,and develop antigen recombinant protein ELISA method against Theileria annulata in cattle,to determine antigen coating concentration on the square ex-periment by using antigen recombinant protein TaSP-Tams1-SPAG1 against Theileria annulata expressed by eukarya as ELISA coating antigen.The results indicated that the optimal concentration of coating anti-gen was 10 μg/mL and the optimal blocking dilution of serum sample was 1 ∶100 in the cross assay.The critical value reacted by ELISA positive was an OD 492nm ≥ 0.348.The variation coefficient of intra-batch and the inter-batch in the repeating tests was less than 12%.No cross reactions were found among the piro-plasms.The improved indirect TaSP-Tams1-SPAG1 ELISA Was highly sensitive specific and reproducible, which provided a technical basis for accurate detection and control of Theileria annulata diseases in cattle.