杂交瘤细胞凝集试验诊断血吸虫病的实验研究

目的用杂交瘤细胞凝集试验(hybridomacellsagglutinationtest,HCAT)诊断实验日本血吸虫病,并探讨杂交瘤细胞在特异性血清中凝集的机制。方法分泌抗血吸虫31/32kDa抗原单抗的杂交瘤细胞H226经特定处理后分别与感染日本血吸虫尾蚴10、30和50条的小鼠血清孵育,动态观察感染后不同时间的凝集反应。上述杂交瘤细胞涂片后与日本血吸虫感染兔血清进行间接免疫荧光试验(IFT),以探讨凝集机制。结果杂交瘤细胞在感染鼠血清中呈现凝集反应:感染后2wk时,50条尾蚴感染组中70％HCAT阳性,至第5周,全部感染小鼠均出现阳性反应。抗原滴度在感染后逐步升高,感染后6wk时达到最高。重度感染鼠的抗原滴度明显高于中、轻度感染鼠。IFT显示,在杂交瘤细胞膜表面呈现特异性的黄绿色荧光,而细胞内未见显色。结论HCAT是一种血吸虫病诊断新方法。

Experimental study on Hybridoma Cells Agglutination Test for Diagnosis of Schistosomiasis Japonica in Mice

Objective To detect the infection of Schistosoma japonicum in mice with a novel test based on agglutination of hybridoma cells and to study the mechanism of the hybridoma cells agglutination. Methods The procedure was developed with a murine cell line H226 producing a monoclonal antibody specific to schistosome 31/32 kDa antigen and sera collected from mice infected with different numbers (10,30,50) of S. japonicum cercariae in different period. Immunofluorescent test was carried out with the hybridoma cells and schistosome-infected sera. Results The circulating antigen was detected by the test as early as 2 weeks after a heavy infection and all mice showed positive results in the test by 5 weeks after infection. The titers of antigen rose along with the lime post infection, and the tilers of sera from heavy infection were statistically higher than that from the mice receiving a lower number of cercariae. Specific yellowish green fluorescence appeared on the membrane of the hybridoma cells; no signal was detected inside. Conclusion Hybridoma cell agglutination test (HCAT) may become useful to diagnose schistosomiasis.