胶体金免疫层析法检测广州管圆线虫感染小鼠血清特异性抗体的研究  

Development of a colloidal gold immunochromatographic assay to measure serum antibodies against Angiostrongylus cantonensis in mice

作　　者：郝振华[1] 黄慧聪[1] 谭峰[1] 梁韶晖[1] 

机构地区：[1]温州医科大学寄生虫学教研室,浙江温州325035

出　　处：《中国病原生物学杂志》2013年第11期1008-1010,共3页Journal of Pathogen Biology

基　　金：浙江省大学生科技创新活动计划(新苗人才计划)项目(No.2011R413038)

摘　　要：目的建立一种无需仪器、操作简单,具较好敏感性和特异性的适用于基层快速检测广州管圆线虫感染的胶体金免疫层析方法。方法将重组广州管圆线虫五期幼虫抗原Ac11蛋白为检测点膜抗原,羊抗兔IgG喷涂在质控线,与胶体金标记兔抗小鼠单抗组装成广州管圆线虫胶体金免疫层析法检测试剂条,用于检测阳性小鼠血清特异性抗体。试验设阴性小鼠血清为对照。结果确定试剂条的检测条件为:检测线重组抗原Ac11用量为4mg/ml(喷量1μl/cm),质控线羊抗兔IgG用量为1.87mg/ml(喷量1μl/cm),胶体金标记兔抗小鼠单抗采用吸光度(A)值为5的喷涂量(喷量2μl/cm)。用该方法检测小鼠血清抗广州管线虫抗体,其敏感性为100%(10/10),特异性为100%(10/10)。结论胶体金免疫层析法检测广州管圆线虫抗体具有较好的敏感性和特异性,经进一步完善后有望用于广州管圆线虫病的临床快速筛查和流行病学调查。Objective To develop a rapid, sensitive, and specific method of measuring antibodies against Angiostrongy, lus cantonensis. Methods A colloidal gold immunochromatographic assay was devised. The recombinant antigen Acl 1 from fifth-stage larvae served as the detecting antigen, goat anti-rabbit IgG-HRP was used to provide the control line, and rabbit anti-mouse monoclonal antibody was labeled with colloidal gold. This assay was used to detect serum-specific anti bodies in infected mice. Serum from uninfected mice served as the control. Results Optimal conditions were as follows= 4 mg/ml （spray volume： 1 μl/cm） recombinant antigen Acll as the detecting antigen, 1.87 mg/ml （spray volume： 1 μl/ cm） goat anti-rabbit IgG-HRP as the control, and absorbance of rabbit anti mouse monoclonal antibody labeled with col loidal gold of 5 （spray volume： 2 μl/cm）. The assay detected antibodies against Angiostrongylus cantonensis in mice scr um with a sensitivity of 100% （10/10） and specificity of 100% （10/10）. Conclusion The colloidal gold immunochron~ atographic assay thus devised had excellent sensitivity and specificity at detecting antibodies against A. cantone^zsis. This assay could be used for clinical diagnosis and epidemiological study of A. cantonensis.

关 键 词：胶体金免疫层析 广州管圆线虫 抗体 

分 类 号：R383.1[医药卫生—医学寄生虫学]