摘要
目的研制一种检测弓形虫病IgG抗体的快速胶体染料试纸条法(DDIA),与国外进口试剂盒进行比较,以评价其临床应用价值。方法以本实验室筛选的胶体染料D1标记羊抗人IgG,以此标记物与弓形虫病人血清中的抗弓形虫IgG抗体结合,用点有弓形虫可溶性抗原的硝酸纤维膜通过层析捕获染料标记羊抗人IgG与弓形虫IgG抗体的复合物,采用正交试验确定DDIA的最佳检测条件;同时对弓形虫病流行病学筛查时的88份血清进行检测并与进口试剂盒的检测结果进行比较。结果DDIA的最佳检测条件为弓形虫可溶性抗原的点膜浓度为1mgml,人IgG点膜浓度为2mgml,血清用量为10μl和羊抗人IgG胶体染料检测液作1∶4稀释时的检测带清晰且背景较浅;DDIA与进口试剂盒的总符合率为97.7%,其中阳性和阴性符合率分别为100%(5252)和94.4%(3436)。结论DDIA与进口试剂盒有较高的符合率,表明该法具有较好的弓形虫病诊断价值,可进一步推广应用。
Objective To establish a rapid, immunodiagnostic assay for the diagnosis of toxoplasmosis immunoglobulin G (IgG). Methods Goat anti-human IgG was conjugated with a blue colloidal dye (D-1) produced in China and used to detect IgG of Toxoplasma gondii in toxoplasmosis patients. The labeled second antibody-antibody complex was captured by soluble antigen of Toxoplasma onto a nitrocellulose membrane dipstick by means of immunochromagraphy. The optimal detecting condition of DDIA was determined by orthogonal experiment. A total of 88 human sera samples obtained in epidemiological investigation of toxoplasmosis were tested with DDIA, its results were compared with that tested by SERION ELISA classic Toxoplasma gondii IgG kit produced from Germany. Results The optimal detection condition of DDIA was that the concentration of soluble antigen of Toxoplasma was 1mg/ml, the concentration of human IgG was 2mg/ml, the detective volume of human serum was 10μl; the total correspondence rate, the positive correspondence rate and negative correspondence rate of DDIA was 97.7%(86/88), 100%(52/52) and 94.4%(34/36) respectively compared with imported IgG-ELISA kit of Toxoplasmosis. Conclusion DDIA has good correspondence rate with imported IgG-ELISA kit of Toxoplasmosis. So DDIA has potential value for immunodiagnosis of toxoplasmosis.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第12期1000-1003,共4页
Chinese Journal of Microbiology and Immunology
基金
江苏省卫生厅重大科技项目(H2006)
