新型自身抗体联合检测斑点免疫金渗滤法的建立及应用评价  

Establishment and evaluation of a novel dot immunogold filtration assay for simultaneous detection of multiple autoantibodies

作　　者：杨湘越[1] 兰小鹏[1] 徐秀云[1] 张惠菊[1] 

机构地区：[1]南京军区福州总医院全军检验医学研究所,350025

出　　处：《实用医学杂志》2009年第7期1147-1149,共3页The Journal of Practical Medicine

基　　金：福建省青年人才科技创新基金资助项目(编号:2002J060)

摘　　要：目的:建立一种简便快速的联合检测自身抗体的斑点免疫金渗滤法(DIGFA)。方法:将多种人自身抗原SmD1、SSA、SSB、Jo-1,采用毕赤酵母重组表达后,纯化抗原点于硝酸纤维素膜上,用来检测血清中相应多种自身抗体,通过胶体金标记的金黄色葡萄球菌A蛋白直接显色,阳性者出现红色斑点。结果:DIGFA检测185份经欧蒙ENA酶斑点法检测为不同自身免疫病患者血清和50份正常人血清,阳性符合率分别为抗SmD191.2%,抗SSA92.8%,抗SSB96.9%,抗Jo-192.6%,阴性符合率为100%(50/50)。与欧蒙ENA酶斑点法对比检测的总符合率为93.0%(172/185),差异无显著性(P>0.05)。结论:DIGFA法简便、快速、准确,可用于临床自身免疫病的联合诊断。Objective To establish a new simple rapid dot immunogold filtration assay （DIGFA） for simultaneous detection of multiple autoantibodies. Methods Autoantigens （SmD1, SSA, SSB, and Jo-1） which were recombined and expressed by yeast were dripped on nitrocellulose membrane （NC）, then were stained by staphylococcal protein A （SPA） marked with colloidal gold. Red dots would appear when the results were positive. Results Sera collected from 185 patients with different autoimmune diseases and 50 healthy volunteers were detected by DIGFA and extractable nuclear antigens dot-enzyme immunoassay （ENAs dot-EIA）. The positive coincidence rates for anti-StuD1, anti-SSA, anti-SSB, and anti-Jo-1 were 91.2% （52/57）, 92.8% （64/69）, 96.9% （31/32）, and 92.6% （25/27）, respectively, while the negative coincidence rates were 100% （50/50）. The total coincidence rate of the results of simultaneous determination of sera for autoantibodies by DIGFA and ENAs dot-EIA was 93.0% （172/185） （P 〉 0.05）. Conclusion DIGFA is a simple, rapid, and accurate method for detecting multiple autoantibodies, which can be applied in clinical simultaneous diagnosis of autoimmune diseases.

关 键 词：自身免疫疾病 自身抗体 斑点免疫金渗滤法 联合检测 

分 类 号：R446.6[医药卫生—诊断学]