抗Sj14-3-3特异性IgY的制备及其诊断日本血吸虫病循环抗原的价值  

Preparation of specific yolk immunoglobulin(IgY) of Anti-Sj14-3-3 in serological diagnosis of Schistosomiasis japonica

作　　者：罗庆礼[1] 周银娣[1,2] 沈继龙[1] 

机构地区：[1]安徽病原生物学省级实验室和人兽共患病安徽省重点实验室 安徽医科大学病原生物学教研室,合肥230032 [2]合肥市第一人民医院检验科,合肥230032

出　　处：《安徽医科大学学报》2012年第9期1011-1014,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家高技术研究发展计划资助项目(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。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 cap- ture 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.

关 键 词：日本血吸虫病 循环抗原 14-3-3蛋白 卵黄抗体 

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