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目的 进一步证明华支睾吸虫成虫 15 / 16 ku蛋白 (Cs15 / 16 )用于诊断的免疫活性 ,同时测定其部分氨基酸顺序 ,便于今后对此蛋白的人工合成等分子生物学研究及其相关免疫学研究。 方法 用高效液相色谱技术 (HPL C)自华支睾吸虫粗抗原 (Clonorchis sinensis adultworm antigens,CAA)中提纯 15 / 16 ku蛋白 ,用酶联免疫吸附实验 (EL ISA)测定其抗原活性 ,并使用蛋白质自动测序仪测定其 N端部分氨基酸顺序。 结果 3个具有抗原活性的纯化样品 (H3、H11及H13)中 ,H13的诊断效果接近 CAA,敏感性达 90 % ,特异性 95 %。分别测获 3个纯样的 N端 15、14和 2 0个氨基酸残基 (aa) ,其中 H11的 N端 14个 aa与 H13的前 14个完全相同。 结论 进一步证明了 Cs15 / 16为有效的诊断抗原 ,测获的部分氨基酸顺序可为更深入的研究提供参考。
Objective To further confirm the immunogenicity of 15/16 ku protein (Cs15 / 16) of adult Clonanthra sinensis for diagnosis and to determine its partial amino acid sequence in order to facilitate the future molecular biology studies of artificial synthesis of this protein and its related immunology the study. Methods 15/16 ku protein was purified from Clonorchis sinensis adultworm antigens (CAA) by high performance liquid chromatography (HPL C), and its antigenic activity was determined by enzyme-linked immunosorbent assay (ELISA) The N-terminal partial amino acid sequence was determined using a protein automated sequencer. Results Among the three purified samples with antigenic activity (H3, H11 and H13), the diagnostic value of H13 was close to that of CAA with a sensitivity of 90% and a specificity of 95%. The N-terminal 15,14 and 20 amino acid residues (aa) of 3 pure samples were detected respectively. Among them, the 14 aa of the N-terminal of H11 were exactly the same as the first 14 of H13. The conclusion further proved that Cs15 / 16 is a valid diagnostic antigen, and the partial amino acid sequence obtained can provide reference for further research.