金标渗滤法测定囊虫特异性抗体试剂盒的研制

Development of Dot Immunogold Filtration Assay Kit for Rapid Detection of Specific Antibodies Against Cysticercus cellulosae

目的研制简易、快速、准确的囊虫病诊断试剂盒,并对其诊断价值进行考核评价。方法以猪囊尾蚴的囊液粗提液为检测用抗原,以胶体金标记的金黄色葡萄球菌蛋白A(SPA)为探针,采用自行设计的渗滤装置制备金标渗滤法(DIGEA)检测试剂盒。检测不同病人血清中囊虫特异性IgG抗体,并以囊虫IgG抗体 ELISA检测试剂盒考核其敏感性和特异性。结果用金标试剂盒检测58人份囊虫病患者血清和100人份健康者血清,其敏感性为98．3％(57/58);特异性为100％(100/100)。用该试剂盒检测25人份包虫病患者血清,6份呈弱阳性,交叉反应率为24％;检测30人份血吸虫病人血清,10人份肺吸虫病人血清,10人份肠道线虫病人血清,均无交叉反应。与ELISA试剂盒检测结果比较,符合率为98．7％,无显著性差异(X2=0．0545,P>0．05)。结论金标渗滤法快速检测囊虫特异性抗体试剂盒敏感性高、特异性强,可与ELISA法相媲美,且操作更为简便、快速,结果判读容易,特别适合临床检测和现场查病。

Objective To develope a simple, rapid, and sensitive diagnostic kit for diagnosis of cysticercosis. Methods Crude extracts of cyst fluid of Cysticercus cellulosae were used as the antigen and SPA was conjugated with colloid gold as the detecting probe. A special pad for dot immunogold filtration assay （DIGFA） was designed according to the reaction procedure. The specific antibodies were detected in different patient sera using DIGFA kit, and Cyt-ELISA kit was utilized in parallel with this kit. Results Sensitivity and specificity of this kit were 98.3%（57/58） and 100% （ 100/100）, respectively. Twenty four percent of cross reaction was found with sera of echinococcosis patients. None cross reaction showed with other helminthiasis such as schistosomiasis, paragonimiasis and intestinal nematode infection. There is not significant difference in comparison with Cyst-ELISA kit （χ^2=0.0545 ,P 〉0.05）. Conclusion This rapid diagnostic kit for detection of specific IgG is rapid, simple and with high sensitivity and specificity. It is very useful for clinical diagnosis and field investigation of Cysticercosis cellulosae.