摘要
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 confirm further the immunological activity of Clonorchis sinensis adultworm 15/16 ku protein (Cs15/16) and to assay its partial amino acid sequence to facilitate the later molecular biological studies on this protein, such as the artificial synthesis of it, and some related immunological studies. Methods We purified 15/16 ku proteins from C. sinensis adultworm antigens (CAA) with high performance liquid chromatography (HPLC), tested their antigenic activities with ELISA and assayed their N end partial aminoacid sequences on a protein auto sequencer. Results Of three 15/16 ku purified samples that had antigenic activity (H3, H11 and H13), H13, with the sensitivity of 90% and the specificity of 95%, had very close effect to that of CAA. 15, 14 and 20 aminoacid remnants (AAs) of three samples, were determined respectively. The 14 AAs of H11 were identical to the 14 N end initial AAs of H13, but had no similarity to that of H3. Conclusion This study further proves that Cs15/16 is effective diagnostic antigen, and the assayed partial aminoacid sequences can provide references for deeper researches in this field.
出处
《中国寄生虫病防治杂志》
CSCD
2001年第4期280-282,共3页
Chinese Journal of Parasitic Disease Control
基金
广东省医学科学技术研究基金资助课题(No.A1 9970 59)~~
