Objective To detect the diagnostic value of using yolk immunoglobulin ( IgY ) and establishing sandwich ELISA in Schistosoma japonicum. Methods IgY and IgG antibodies were prepared and purified from the yolk and the sera of rabbits immunized with rSj4-3-3. Double antibody sandwich ELISA using the specific IgG for capture and IgY for detection was set up to test sera of 72 cases of acute and 128 cases of chronic schistosomiasis. 132 sera of normal individuals were used as control. Additionally, 42 patients with paragonimiasis, 60 with clonorchiasis and 46 with hookworm infections were subjected to cross-reaction investigation. Results The positive rates of acute and chronics schistosomiasis were 73. 6% and 48. 4% when IgY was used as detection antibody. The negative rate of normal human sera was 94. 7% ; the cross-reactions with paragonimiasis , clonorchiasis, and hookworm infections were found to be 7. 1% , 6. 7% , and 6. 5% , respectively. Conclusion IgY based double antibody sandwich ELISA used for the detection of circulating antigens has a potential and practical value in the diagnosis of schistosomiasis.%目的 制备抗重组日本血吸虫信号转导蛋白14-3-3(Sj14-3-3)的鸡卵黄免疫球蛋白(IgY),建立双抗体夹心ELISA诊断体系(IgY-ELISA),探讨用于日本血吸虫病的诊断价值.方法 利用Sj14-3-3制备并纯化卵黄抗体IgY和兔多克隆IgG抗体;以IgG为捕获抗体,IgY为检测抗体建立双抗体夹心ELISA诊断方法,检测72例急性血吸虫病患者、128例慢性血吸虫病患者、132例正常人、42例肺吸虫患者、60例华支睾吸虫患者和46例钩虫感染者血清.探讨基于IgY的循环抗原检测的应用价值.结果 成功制备并鉴定了抗Sj14-3-3特异性IgY和IgG抗体,建立了双抗体夹心ELISA检测体系;检测急性血吸虫病患者和慢性血吸虫病患者血清的阳性率分别为73.6 %和48.4 %,正常人血清的阴性率为94.7 %;与肺吸虫患者、华支睾吸虫患者、钩虫感染者血清的交叉反应分别为7.1%、6.7%、6.5%.结论 建立了基于IgY的双抗体夹心ELISA检测体系,用于日本血吸虫病患者血清的检测具有较高的敏感性和特异性,可用于日本血吸虫病的辅助诊断.
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