基于IgY抗体检测日本血吸虫循环抗原的间接红细胞凝集试验的建立  

Establishment of chicken egg yolk antibodies （IgY） -based indirect hemagglutination test for detecting circulating antigens of Schistosoma japonicum

作　　者：蔡玉春[1] 郭俭[1] 陈韶红[1] 田利光[1] 陈木新[1] 艾琳[1] 张玲玲[1] 陈家旭[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾、血吸虫病和丝虫病合作中心,上海200025

出　　处：《国际医学寄生虫病杂志》2012年第1期5-8,共4页International JOurnal of Medical Parasitic Diseases

基　　金：国家科技支撑计划（2008BA156803）;国家科技重大专项（2008ZX10004-011）

摘　　要：目的建立基于I“的血吸虫循环抗原的间接红细胞凝集试验（indirect hemagglutination test，IHA）并初步探讨IgY在诊断日本血吸虫病中的应用价值。方法以日本血吸虫卵可溶性虫抗原（soluble egg antigen，SEA）免疫海蓝蛋鸡，获得抗SEA的IgY抗体。取人O型红细胞醛化、鞣化，形成鞣化后红细胞，并分别用磷酸盐、碳酸盐和柠檬酸盐缓冲液稀释抗SEA-IgY抗体，以确定IgY致敏红细胞最佳缓冲液。将IgY溶解于最佳的缓冲液中致敏鞣化后红细胞，形成IgY红细胞检测液，用此检测液分别检测感染30、20、10条日本血吸虫尾蚴和正常小鼠血清各10份，初步检测不同感染度小鼠血清的循环抗原。结果pH7．2磷酸盐缓冲液溶解的IgY所得致敏红细胞检测血清最佳，效价可达1：320。用此IgY—IHA法检测中、高感染度血清（感染20条和30条日本血吸虫尾蚴）阳性率为100％，低感染度血清（感染10条日本血吸虫尾蚴）阳性率为70％，对照组10只小鼠血清全为阴性，特异性为100％。结论建立了IgY-IHA法，此法检测日本血吸虫病循环抗原具有较高的敏感性。Objective To establish anti-SEA-IgY based indirect hemagglutination test （IHA） method of detecting circulating antigen of Schistosoma japonicum and explore its application value for diagnosis of schistosomiasis. Methods Hens were immunized with soluble egg antigen （SEA） of S. japonicum, specific anti-SEA IgY was purified from eggs after immunization. The anti-SEA IgY was diluted with three different buffer as phosphate （ PBS, pHT. 2）, carbonate （ pH 9.6） and citrate buffer to choose the best buffer. After aldehyding and tanning, the human O type erythrocytes were sensitized with the anti-SEA IgY which was dissolved into the best buffer, formatting the IgY adsorbed erythrocytes test solution. With this test solution, 30 sera of mice infected with 10,20 or 30 S. japonicum cercariae respectively and 10 sera of normal control mice were tested for detecting circulating antigens in the sera of mice infected with S. japonicum. Results Erythrocytes which were sensitized with the anti-SEA IgY dissolving into phosphate buffer （pH7.2） gave the best results, the best titer was up to 1： 320. With this IgY-IHA method, the positive rate of sera of high degree infection（mice infected with 20 and 30 cercariae） were 100%, while the positive rate of sera of low degree infection（ mice infected with 10 cercariae ） were 70%, 10 sera of control group mice were all negative. Conclusion The established IgY-IHA method exhibited higher specificity and＇sensitivity in detecting circulating antigen of S. japonicum.

关 键 词：日本血吸虫 循环抗原 IGY 间接红细胞凝集试验 

分 类 号：R532.14[医药卫生—内科学]