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  

Immunodiagnosis-associated antigens of Pagumogonimus skrjabini adult worm with immunohistochemical technique

作　　者：牛靖萱[1] 张锡林[2] 王利芳[1] 陈文碧[1] 何谐[2] 陈琳[2] 

机构地区：[1]泸州医学院病原生物学教研室,四川泸州646000 [2]第三军医大学基础医学部病原生物学教研室,重庆400038

出　　处：《第三军医大学学报》2010年第7期654-657,共4页Journal of Third Military Medical University

基　　金：四川省教育厅科研基金(2005A067);日本第二期健康科学基金(JPH20070512)~~

摘　　要：目的初步分析斯氏狸殖吸虫成虫表膜和肠相关抗原,探讨肺吸虫病的免疫诊断的特异性诊断抗原。方法斯氏狸殖吸虫成虫制备成15μm的冰冻切片,肺吸虫患者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。结果肺吸虫患者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。结论肺吸虫患者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。Objective To investigate tegument antigens and intestine-associated antigens of Pagumo-gonimus skrjabini（P.skrjabini）adult worm and provide the immunodiagnostic antigen for clinical diagnosis.MethodsThe adults of P.skrjabini were sectioned at a thickness of 15 μm.Serum specific antigens from patients with P.skrjabini infection was used for immunological study.The immunodiagnosis-associated antigens were identified by using immunogold techniques,immune electron microscopy,enzyme-labeled immunoassay and laser scanning confocal microscopy.ResultsBoth immunogold techniques and enzyme-labeled immunoassay showed that specific antigens were concentrated in the tegument and enteric cavity.Confocal laser scanning microscopy showed that semi-quantitative analysis of tegument antigen of average gray value was 80.65,and that of intestine antigen was 71.25.At the ultrastructural level,the immunogold labeling showed deposit of gold particles on the surface membrane of tegument and enteric cavity.ConclusionIdentification of adult specifical antigens of P.skrjabini by using paragonimiasis sera are tegument-and intestine-associated antigens,therefore,immunodiagnostics of paragonimiasis are the adult components of the surface membrane and intestine.

关 键 词：斯氏狸殖吸虫 表膜 肠腔 免疫组化 

分 类 号：R383.2[医药卫生—医学寄生虫学] R329.11[医药卫生—基础医学]