免疫组化技术对斯氏狸殖吸虫成虫免疫诊断相关抗原的初步研究.docVIP

免疫组化技术对斯氏狸殖吸虫成虫免疫诊断相关抗原的初步研究 牛靖萱1，张锡林2，王利芳1，陈文碧1，何谐2，陈琳2 （1．泸州医学院病原生物学教研室，泸州646000；2.第三军医大学病原生物学教研室，重庆400038） 摘要：目的 通过对斯氏狸殖吸虫成虫表膜和肠相关抗原的初步研究，为临床肺吸虫病的免疫诊断提供特异性诊断抗原。方法 斯氏狸殖吸虫成虫制备成15μm的冰冻切片, 肺吸虫病人血清识别斯氏狸殖吸虫成虫特异性抗原，采用免疫金标记和免疫酶标记检测技术对抗原进行定位，激光共聚焦显微镜技术进行半定量分析以及免疫透射电镜对其细微结构的观察。分实验组和对照组。结果 肺吸虫病人血清识别的斯氏狸殖吸虫成虫抗原定位于虫体的表膜和肠腔，表膜抗原半定量分析平均灰度值为80.65，肠腔抗原半定量分析平均灰度值为71.26，电镜下可见胶体金附着于肠腔和表膜的双层结构。结论 肺吸虫病人血清识别的斯氏狸殖吸虫成虫抗原都为表膜相关抗原和肠相关抗原，因此与肺吸虫病免疫诊断相关的诊断抗原为虫体表膜和肠上皮产生的相关蛋白。 关键词：斯氏狸殖吸虫； 表膜 ；肠腔 ；免疫组化 Primary study on immunodiagnosis-associated antigens of Pagumogonimus skrjabini adult with immunohistochemisty technique Niu Jing-xuan1 ,Zhang Xin-lin2 ,Wang Li-fang1 ,Chen Wen-bi1 ,He Xie2 , Che Lin2 (1 Luzhou Medical College Department of pathogenic biological,SiChuan 646000,China；2 Third Military Medical University Department of Biology pathogen，ChongQing 400038,China) Abstract： Objective To research tegument antigens and intestine-associated antigens of Pagumogonimus skrjabini adult worm and provide the immunodiagnostic antigen for clinical diagnosis. Methods. The adlut of P.skrjabini were sectioned at a thickness of 15 μm，and used to localize specific antigen by the sera infected by Pagumogonimus skrjabini. The immunodiagnosis-associated antigens were identified by using immonogold techniques, immune electron microscopy, enzyme-labeled immunoassay and laser scanning confocal microscopy techniques.. Results Both immonogold techniques and enzyme-labeled immunoassay showed that specific antgen was concentrated in the tegumental and intestine. Confocal laser scanning microscope showed that semi-quantitative analysis of tegument antigen of average gray value is 80.65 and semi-quantitative analysis of intestine antigen of average gray value is 71.25. At the ultrastructural level ,the immunogold labeling showed deposit of gold particles on the tegumental and on the surface membrane and intestine. Conclusion Identification adult specifical antigens of Pagumogonimus skrjabini by using