斑点金免疫渗滤法联合检测糖尿病自身抗体的研究  

Development of dot immunogold filtration assay for the simultaneous detection of diabetes autoantibodies

作　　者：李涛[1] 程立庆[1] 陈兵[1] 

机构地区：[1]第三军医大学西南医院内分泌科,重庆400038

出　　处：《免疫学杂志》2014年第1期70-74,共5页Immunological Journal

基　　金：第三军医大学科技成果转化基金(2010XZH07)

摘　　要：目的构建联合检测糖尿病自身抗体的斑点金免疫渗滤法(dot immunogold filtration assay,DIGFA)。方法将重组人胰岛素、谷氨酸脱羧酶、蛋白酪氨酸磷酸酶和锌转运体8抗原点样于硝酸纤维素膜上,制备胶体金标记链霉亲和素作为显色探针,建立斑点金免疫渗滤法检测1型糖尿病(type 1 diabetes,T1DM)患者外周血中相应的自身抗体,并对方法性能开展评估。结果DIGFA法检测42例T1DM敏感性66.7%,检测100例健康对照特异性97%。与ELISA法相比对T1DM自身抗体检出率无显著差异(P>0.05)。3个批次的免疫渗滤装置检测结果一致,4℃保存10周内检测性能稳定。结论本研究构建的斑点金免疫渗滤法准确、快速、简便,为联合检测糖尿病自身抗体提供了一种高效的新途径。This study designed to develop a dot immunogold filtration assay （DIGFA） for simuhaneous detection of multiple diabetes autoantibodies. Recombinant human insulin, glutamic acid decarboxylase （GAD）, tyrosine phosphatase 2 （IA-2）, and zinc transporter 8 （ZnT8） antigen were coated onto nitrocellulose membrane, and streptavidin was conjugated with colloid gold as stained probe. The dot immunogold filtration assay was established and used to detect autoantibodies in sera of type 1 diabetes. Test result showed that the sensitivity of DIGFA was 66.7% in 42 T1DM patients and the specificity was 97% in 100 healthy individuals; there was no significant difference in the detecting results of T1DM patients between DIGFA and ELISA kit （P〉 0. 05）. Furthermore, test results of 3 batches of immunogold filtration device were consistent and stable performance was exhibited after 10 weeks later stored under 4 ℃. In conclusion, the DIGFA is an accurate, rapid, and simple method which can provide a novel efficient approach to simultaneous detection of multiple diabetes autoantibodies.

关 键 词：1型糖尿病 自身抗体 斑点金免疫渗滤试验 

分 类 号：R446.6[医药卫生—诊断学]