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目的：制备抗隐孢子虫单克隆抗体（ＭｃＡｂ）并对其特异性进行鉴定。方法：用纯化的人源微小隐孢子虫卵囊免疫ＢＡＬＢ／ｃ小鼠，杂交瘤技术制备ＭｃＡｂ，间接免疫荧光（ＩＦＡ）和免疫印迹试验分析其特性。结果：制备出Ｚ４Ｃ８、Ｚ２Ｂ６、Ｚ３Ｄ７和Ｚ３Ｄ２四株能稳定分泌抗隐孢子虫ＭｃＡｂ的杂交瘤细胞株。经鉴定，Ｚ４Ｃ８和Ｚ３Ｄ７为ＩｇＭ，Ｚ３Ｄ２和Ｚ２Ｂ６为ＩｇＧ１，轻链均为κ链，与多种肠道病原体及肺孢子虫无交叉反应。除Ｚ２Ｂ６在间接免疫荧光试验中识别卵囊壁外，其余均识别隐孢子虫子孢子（ＣＳＰ）。免疫印迹定位表明，四株ＭｃＡｂ分别识别４２条ＣＳＰ抗原中的２０．５ｋＤａ、６０．５ｋＤａ、３３ｋＤａ和９５ｋＤａ４个条带，其中Ｚ４Ｃ８和Ｚ３Ｄ７均识别２０．５ｋＤａ主要抗原带。结论：这四株ＭｃＡｂ在识别ＣＳＰ抗原表位上具有不同的特性，但对隐孢子虫抗原均呈高度特异性反应。
Objective: To prepare anti-Cryptosporidium monoclonal antibody (McAb) and to identify its specificity. Methods: BALB / c mice were immunized with the purified human Crytosporidium parvum oocysts, hybridomas were used to prepare McAbs, indirect immunofluorescence (IFA) and Western blotting were used to analyze their characteristics. Results: Four strains of hybridoma cell lines secreting anti-Cryptosporidium McAb were prepared by four strains of Z4C8, Z2B6, Z3D7 and Z3D2. It has been identified that Z4C8 and Z3D7 are IgM, Z3D2 and Z2B6 are IgG1, both light chain are kappa chain, and no cross-reaction with various intestinal pathogens and Pneumocystis. Except that Z2B6 recognized the oocyst wall in the indirect immunofluorescence assay, all others recognized Cryptosporidium sporozoites (CSP). Immunoblotting showed that the four McAbs recognized four bands of 20.5kDa, 60.5kDa, 33kDa and 95kDa respectively from 42 CSP antigens, of which Z4C8 and Z3D7 all recognized the major antigenic band of 20.5kDa. CONCLUSION: These four McAbs have different characteristics in recognizing CSP epitopes, but have highly specific response to Cryptosporidium antigens.