基于IgY的ELISA用于日本血吸虫病循环抗原的检测  

Detection of circulating antigen of Schistosoma japonicum using yolk immunoglobulin-based ELISA

作　　者：周银娣[1] 罗庆礼[1] 余轶婧[1] 张诗海[1] 李静[1] 沈继龙[1] 

机构地区：[1]安徽医科大学病原生物学教研室,人畜共患病安徽省重点实验室,重要遗传病基因资源利用教育部省部共建重点实验室,合肥230032

出　　处：《安徽医科大学学报》2008年第4期372-376,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家高技术研究发展计划资助项目("863"计划;编号:2006AA02A251);安徽高校省级自然科学研究计划项目(编号:KJ2008B285)

摘　　要：目的以鸡卵黄免疫球蛋白(IgY)替代哺乳动物来源的IgG抗体,建立双抗体夹心ELISA用于日本血吸虫病的诊断。方法制备并纯化抗日本血吸虫可溶性虫卵抗原(SEA)的卵黄抗体IgY和抗SEA的兔多克隆IgG抗体;以抗SEA的IgG为捕获抗体,IgY为检测抗体的双抗体夹心ELISA诊断方法,共检测血清443份,其中急性血吸虫病患者血清68例,慢性血吸虫病患者180例,正常人100例,肺吸虫感染者20例,华支睾吸虫感染者48例和钩虫感染者血清27例。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79.4%和64.4%,最低灵敏度为9.3μg/L;正常人血清的阴性率为92%;与肺吸虫、华支睾吸虫、钩虫感染者血清的交叉反应分别为20%、20.8%、14.8%。结论建立了基于IgY的双抗夹心ELISA检测体系,用于日本血吸虫SEA的检测具有一定特异性和较高的敏感性,可试用于日本血吸虫病的辅助诊断。Objective To use yolk immunoglobulin （ IgY ） , instead of IgG antibodies generated from mammalian animals, and establish sandwich ELISA to detect the circulating soluble egg antigens （SEA） of Schistosoma japonicum for the diagnosis of schistosomiasis. Methods IgY and IgG antibodies against SEA from the yolk and the sera of rabbits immunized with SEA were prepared and purified first. Then double antibody sandwich ELISA were developed using the specific IgG as capture antibody and the IgY as detection antibody. Following that 443 sera cases were tested using the new method developed, including 68 cases of acute and 180 cases of chronic schistosomiasis. In addition, 100 sera specimens of normal individuals were used as control and 20 patients with paragonimiasis, 48 with clonorchiasis and 27 with hookworm infections were used to perform cross-reaction investigation. Results The positive rates of acute and chronics schistosomiasis were 79. 4% and 64. 4% respectively when IgY was used as detection antibody. The sensitivity was 9. 3 μg/L, and specificity of normal human sera was 92% ; the cross-reaction with paragonimiasis, clonorchiasis, and hookworm infections were found to be 20%, 20. 8%, and 14. 8%, respectively. Conclusion IgY antibody used for the detection of circulating antigens has potential and practical value in the diagnosis of schistosomiasis.

关 键 词：血吸虫病 日本/诊断 

分 类 号：R532.210.4[医药卫生—内科学] R341[医药卫生—临床医学]