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为了对华支睾吸虫重组抗原的特性进行鉴定及虫体定位,进而推测其生物学功能,通过对囊蚴与成虫的cDNA表达文库的免疫学筛选,获得若干个强反应原性抗原蛋白,在此基础上,发现2个含有20~30个氨基酸的高度重复序列CsSQ1和CsSQ2,利用基因序列合成多肽,再将多肽与载体蛋白进行偶联,制备重组抗原rCsSQ1和rCsSQ2,免疫动物制备多克隆抗体,利用硫酸铵沉淀法及亲和层析法纯化抗体,最后利用免疫荧光技术对重组抗原进行免疫学虫体定位。结果显示,重组抗原rCsSQ1和rCsSQ2分别定位于华支睾吸虫的口吸盘和腹吸盘。利用免疫荧光技术成功地确定了华支睾吸虫这两种抗原在虫体中的定位,推测它们可能与华支睾吸虫的免疫黏附作用有关,为华支睾吸虫抗原的免疫学特性及其在诊断中的应用研究提供了依据。
In order to identify the characteristics of the recombinant antigen of Clonorchis sinensis and the localization of parasites, and then to speculate on its biological function, we obtained several strong antigenic antigenic proteins through immunological screening of the cDNA expression library of metacercariae and adult worms. On this basis, we found two highly repetitive sequences CsSQ1 and CsSQ2 containing 20-30 amino acids, synthesized the polypeptide by gene sequence, and then conjugated the polypeptide with the carrier protein to prepare recombinant antigens rCsSQ1 and rCsSQ2, and immunized animals to prepare polyclonal antibodies , Using ammonium sulfate precipitation method and affinity chromatography purification of antibodies, and finally using immunofluorescence technology of recombinant antigens for immunological host localization. The results showed that the recombinant antigens rCsSQ1 and rCsSQ2 were located in the sucker and abdominal sucker of Clonorchis sinensis respectively. The immunofluorescence technique was used to successfully determine the localization of the two antigens of Clonorchis sinensis in the parasites. It is speculated that they may be related to the immune adherence of Clonorchis sinensis and the immunological characteristics of Clonorchis sinensis antigen Diagnostic applications provide a basis for research.