摘要
目的 研制一种检测弓形虫病Ig M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D- 1标记兔抗人Ig M,以此标记物与弓形虫病人血清中的抗弓形虫Ig M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兔抗人Ig M与弓形虫Ig M抗体的复合物,采用正交试验确定DDIA的最佳检测条件;对2 5份弓形虫Ig M抗体阳性血清和5 0份正常人血清进行检测;同时对弓形虫病流行病学筛查时的84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1mg/ m l,血清用量为10μl和兔抗人Ig M胶体染料检测液作1∶2 0稀释时的检测带清晰且背景较浅;2 5份弓形虫Ig M抗体阳性血清均为阳性,5 0份正常人血清有3份为Ig M抗体阳性;DDIA与进口试剂盒的总符合率为97.6 % ,其中阳性和阴性符合率分别为10 0 .0 % (35 / 35 )和95 .9% (47/ 4 9)。结论 DDIA简便快速,有较好的敏感性和特异性,同时与进口试剂盒有较高的符合率,表明该法具有较高的弓形虫病诊断价值。
Objective To establish a rapid, simple immunodiagnostic assay for diagnosis of toxoplasmosis immunoglobulin M ( IgM). Methods Rabbit anti-human IgM was conjug a ted with a blue colloidal dye (D-1) produced in China and used to detect antibo d y IgM of Toxoplasma gondii in toxoplasmosis pati ents. The labeled second antibod y-antibody complex was captured by soluble antigen of Toxoplasm a onto a nitrocel lulose membrane dipstick by means of immunochromagraphy. The optimal det ecting c ondition of DDIA was determined by orthogonal experiment. Twenty-five IgM-pos it ive sera and 50 normal sera were detected by DDIA. A total of 84 human sera samp les obtained from epidemiological investigation of toxoplasmosis were tested wi th DDIA, its results were compared with that tested by SERION ELISA classic Toxoplasma gondii IgM kit produced from Germany. Results The optimal detection condition of DDIA wa s that the concentration of solu ble antigen of Toxoplasma was 1 mg/ml, The D-1 dye labeled Rabbit anti-human IgM diluted in 1∶20 has clear detective dot and low background, the detective volum e of human serum was 10 μl. Twenty-five IgM-positive sera samples were positi ve, and there were three IgM-positive sera samples in 50 normal sera. The total cor r espondence rate, the positive correspondence rate and negtive correspondence rat e of DDIA were 97.6%(82/84), 100% (35/35) and 95.9% (47/49) respectively compa red with imported IgM-ELISA kit of toxoplasmosis. Conclusion The DDIA is simple and fast,and has good sensitivity and specificity, it al so has good correspondence rat e with imported IgM-ELISA kit of toxoplasmosis. It indicated that DDIA has pote ntial value for immunodiagnosis of toxoplasmos is.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
2005年第2期93-96,共4页
Chinese Journal of Schistosomiasis Control
基金
江苏省卫生厅重大科技项目 (H2 0 0 6)
