寄生虫与医学昆虫学报2011，Vol.18Issue(4)：193-197,5.DOI:10.3969/j.issn.1005-0507.2011.04.001
抗日本血吸虫SEA特异性IgY应用于循环抗原检测的研究
Detection of Circulating Schistosome Antigen with Anti-SEA Specific Egg Yolk Antibody (IgY)
摘要
Abstract
A sensitive and specific double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect circulating schistosome antigen with chicken egg yolk immunoglobulin antibodies (IgY ) against soluble egg antigen (SEA). Hens were subcutaneously immunized with SEA and the crude IgY was extracted from egg yolk by water dilution method. Purified IgY antibodies and rabbit IgG antibodies against 5. japonicum were labeled with horseradish peroxidaserespectively (IgY-E and IgG-E). 4 double-antibody sandwich ELISA methods were developed using IgY or IgG as capture antibody and using IgY-E or IgG-E as detecting antibody respectively. The detection limits of SEA of 4 ELISA methods were analyzed. Then human serum from patients and healthy people were detected in parallel by established 4 ELISA methods, including 6 serum samples from acute schistosomiasis patients, 29 from chronic schistosomiasis patients, 118 from healthy people, 9 from clonorchiasis patients, 15 from paragonimiasis patients, and 7 from toxoplasmiasis patients. The results showed that the minimal detectable concentration of SEA was 7. 5 ng/ml by IgY based-sandwich ELISA, the sensitivity was higher than IgG based-sandwich ELISA by 4 times. All sandwich ELISA methods showed a sensitivity of 100% for acute schistosomiasis patients, a specificity of 99. 1% for healthy persons; no cross-reaction with Toxoplasmiasis and clonorchiasis; The positive rate of samples from 29 chronic schistosomiasis patients and 15 paragonimiasis patients were 79. 3% , 6. 7% respectively when using IgY as capture antibody, while the positive rate were 75. 8% , 6. 7% and 72. 4% , 13.3% when using IgG as capture antibody, IgY-E and IgG-E as detecting antibody respectively, indicating the assay involved the use of IgY was better than that of rabbit anti-schistosoma IgG. The novel double-antibody sandwich ELISA assay using anti-SEA IgY appears to be sensitive and specific for detection of circulating schistosome antigens. It is the promising assay for immunodiagnosis of S. japonicum infections.关键词
IgY/日本血吸虫/循环抗原/双抗体夹心ELISAKey words
Egg yolk immunoglobulin antibody ( IgY ) / Schistosoma japonicum/ Circulating antigen/ Double-antibody sandwich ELISA引用本文复制引用
楼涤,陆绍红,陈睿,童群波,丁建祖,卓敏敏..抗日本血吸虫SEA特异性IgY应用于循环抗原检测的研究[J].寄生虫与医学昆虫学报,2011,18(4):193-197,5.基金项目
浙江省卫生厅项目(2007A003) （2007A003）
浙江省科技计划项目(2009F20036) （2009F20036）
浙江省医学重点学科群项目(XKQ-009-003) （XKQ-009-003）
浙江省卫生高层次创新人才培养工程项目 （）
