摘要
目的建立基于IgY的双抗体夹心ELISA用于囊尾蚴病的诊断。方法制备并纯化抗囊尾蚴循环抗原(CA)卵黄抗体(IgY),建立以抗CA的IgY为捕获抗体,酶标记抗CA的单克隆抗体1A5为检测抗体的双抗体夹心ELISA法,共检测样品450份,并与捕获抗体和检测抗体均为单克隆抗体的ELISA法比较,验证方法的敏感性、特异性与实用性。结果成功制备并鉴定了特异性IgY抗体,建立了基于IgY的双抗体夹心ELISA检测体系。IgY-ELISA和双单抗-ELISA检测囊尾蚴CA的灵敏度分别为8.3μg/L和13.9μg/L。IgY-ELISA检测囊尾蚴病患者血清与脑脊液的CA阳性率分别为100%(139/139)与89.5%(17/19),囊尾蚴病猪血清的阳性率100%(222/222),健康人与健康猪血清的阴性率为100%。结论建立的基于lgY的双抗体夹心ELISA检测囊尾蚴CA用于囊尾蚴病诊断,具有较高的特异性和敏感性,可用于囊尾蚴病的辅助诊断。
Objective To develop a sensitive and specific double antibody sandwich enzyme-linked immunosorbent assay(ELISA) to detect circulating antigen(CA) of Taeniasoliumcysticercosis with chicken egg yolk immunoglobulin antibodies(IgY).Methods Hens were subcutaneously immunized with CA and the crude IgY was extracted from egg yolk by water dilution method.A sandwich ELISA had been developed by purified IgY antibodies as capture antibody and monoclonal antibodies labeled with peroxidase as detecting antibody.The detection limits of CA were analyzed.The sera and cerebrospinal fluid of patients,the sera of healthy people,sick pigs and healthy pigs were detected in parallel by the established ELISA methods.It's sensitivity and specificity were evaluated by comparison with ELISA based monoclonal antibodies.Results The minimal detectable concentration of CA was 8.3 and 13.9 μg/ml by sandwich ELISA based IgY and monoclonal antibodies,respectively.The positive rates of samples from 139 patients,19 cerebrospinal fluid of patients and 222 sick pigs were 100%(139/139),89.5%(17/19) and 100%(222/222) by sandwich ELISA based IgY respectively.The negative rates of samples from 50 healthy people and 20 healthy pigs were 100%.Conclusion The novel double-antibody sandwich ELISA using anti-CA IgY appears to be sensitive and specific for detection the CA of Taenia solium cysticercosis.It is the promising assay for immunodiagnosis of Taenia solium cysticercosis.
出处
《东南国防医药》
2013年第2期135-137,共3页
Military Medical Journal of Southeast China
关键词
囊尾蚴病
IGY
循环抗原
双抗体夹心酶联免疫吸附实验
Taenia solium cysticercosis
egg yolk immunoglobulin antibody(IgY)
circulating antigen
sandwich ELISA
