光敏生物素标记质粒DNA检测双链DNA抗体ELISA的建立及初步应用

Estabilishment and initial application of photobiotinylated plasmid DNA ELISA for the detection of anti dsDNA

建立一种既可定性也可定量检测血清抗双链DNA抗体的ELISA ,进一步提高对系统性红斑狼疮 (SLE)诊断和监测的水平。以链霉亲合素处理微孔板 ,包被光敏生物素标记的质粒DNA ,用HRP 羊抗人IgG测定血清双链DNA抗体。分别以自制ELISA和Bindingsite公司ELISA抗dsDNA定量试剂盒和胶体金斑点免疫渗滤法定性检测了 32例活动期SLE ,6例类风湿关节炎 (RA) ,30例健康人血清 ,阳性符合率分别为 96 9%和 93 8% ,阴性符合率均为 96 7%。定量检测了 5例SLE ,结果较为一致。

To estabilish a qualitative as well as quantitative ELISA for the detection of anti dsDNA to improve diagnostic and monitoring level for SLE.Photobiotinylated plasmid ds DNA is coated after streptavidin procoated,to detect serum anti dsDNA with HRP sheep anti human IgG.Using a self prepared coated ELISA plate and anti dsDNA quantitative kit of Binding site company of Birmingham medical college U K,and dot immunogold filtration assay have detedted sera of 32 patients with active SLE,6 rheumatoid arthritis and 30 normal controls,results showed,positive consistent rate is respectively 96 9% and 93 8% and negative consistent rate is 96 7%,5 patients with SLE were tested with the detection of quantitative,results were almost consistent.Initial application indicated the ELISA plate achieved good results and worth further studying and expanding detectable serum samples of patients.