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在家兔感染日本血吸虫过程的追踪研究中用间接荧光抗体试验分析日本血吸虫抗原的定位,或许由于成虫、虫卵和尾蚴存在共同抗原,故难于说明感染过程中抗原的期特异性。本研究用不同剂量干重的日本血吸虫雄虫、雌虫、虫卵抗原及肝片吸虫、华支睾吸虫、菲律宾肺吸虫的盐水提取粗制抗原和福氏完全佐剂一起分4次皮内免疫家兔,每周1次,第5次为强化剂,不加佐剂。2周后收集血清,然后通过铵盐沉淀及DEAE纤维素柱层析分离和纯化抗日本血吸虫成虫及虫卵的IgG,并用异硫氰基荧光素标记(即FITC-Ad-IgG和FITC-Egg-IgG)。日本血吸虫成虫、虫卵、尾蚴的组织切片分别用冰丙酮及福尔马林固定以用作荧光试验的抗原。间接荧光法用不同稀释度的免疫兔血清与抗原切片孵育,然后用荧光标记的抗兔γ-球蛋白检测,直接荧光试验用FITC-Ad-IgG及FI-TC-Egg-IgG检测,免疫荧光抑制试验,则用
In the follow-up study of rabbits infected with Schistosoma japonicum, the indirect fluorescent antibody test was used to analyze the localization of Schistosoma japonicum antigens. It may be difficult to explain the stage specificity of the antigens during the infection due to the existence of common antigens in adults, eggs and cercariae. In this study, different doses of dry weight of Schistosoma japonicum, females, egg antigen and Fasciola hepatica, Clonorchis sinensis, Paragonimus philippinarum brine extraction crude antigen and Freund’s complete adjuvant with 4 times intradermal Immunized rabbits, once a week, the fifth for the strengthening agent, without adjuvant. After 2 weeks, the serum was collected, and then the IgG against adult worms and eggs of S. japonicum were isolated and purified by ammonium sulfate precipitation and DEAE cellulose column chromatography, and labeled with isothiocyanine fluorescein (ie FITC-Ad-IgG and FITC-Egg -IgG). Schistosoma japonicum adult worm eggs, cercariae tissue sections were fixed with ice acetone and formalin for fluorescence test antigen. Indirect fluorescence assay was performed with different dilutions of immunized rabbit sera incubated with antigenic sections and then detected with fluorescently labeled anti-rabbit γ-globulin. Direct fluorescence assays were performed with FITC-Ad-IgG and FI-TC-Egg-IgG and immunofluorescence Suppression test, then use