抗Sj14-3-3特异性IgY的制备及其用于日本血吸虫病循环抗原的诊断.docVIP

 抗 Sj14-3-3 特异性 IgY 的制备及其用于日本 血吸虫病循环抗原的诊断# 罗庆礼1，周银娣1,2，沈继龙1** 5 10 15 （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） 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 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，检测急性血吸虫病患者血清 72 例，慢性血吸虫病患者 128 例， 正常人血清 132 例，肺吸虫患者 42 例，华支睾吸虫患者 60 例和钩虫感染者血清 46 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 43.4 %，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。 关键词：血吸虫病；循环抗原；14-3-3 蛋白；卵黄抗体 20 25 30 35 40 中图分类号：R532.210.4; R341; R392.11 Preparation of specific yolk immunoglobulin (IgY) of Anti-Sj14-3-3 to serological diagnosis of Schistosomiasis japonica LUO Qingli, ZHOU Yindi, SHEN Jilong (Provincial Laboratory of Microbiology Parasitology Anhui, Department of Microbiology and Parasitology, Anhui Medical University, Hefei 230032, PR China) Abstract: ITo use yolk immunoglobulin (IgY) instead of IgG antibodies generated from mammalian animals and establish sandwich ELISA to detect the circulating antigens of Schistosoma japonicum for the diagnosis of schistosomiasis. Prepared and purified IgY and IgG antibodies against rSj14-3-3 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 cases, including 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. The positive rates of acute and chronics schistosomiasis were 73.6 % and 48.4 % when IgY was used as detection antibody. The specificity of normal human sera was 94.7 %