Maniatis A K, Yu L, Miao D, Nelson K, Eisenbarth G S
The Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver 80262, USA.
J Autoimmun. 2001 Feb;16(1):71-6. doi: 10.1006/jaut.2000.0457.
The purpose of the current study was to develop and evaluate rapid assays for autoantibodies to GAD65 (GAA), ICA512bdc/IA-2 (ICA512AA), and insulin (microIAA, mIAA) as a potential tool for identification of cadaveric pancreas donors who were at high risk for developing diabetes. The study included 154 new onset diabetic, prediabetic, and healthy control subjects. Subjects were evaluated for all three autoantibodies in three separate assays: (1) standard (std) assay with a 24-h or 72-h incubation at 4 degrees C (combined GAA/ICA512AA or mIAA, respectively), (2) rapid assay with 1-h room temperature (RT) incubation, and (3) rapid assay with 2-h RT incubation. The serum samples from 777 organ donors were also evaluated for all three autoantibodies and all the positive samples from standard assay evaluated with the 1-h incubation assay. Simple linear regression analyses revealed excellent correlation between the standard assay and the rapid assays for all three autoantibodies, as follows: (1) GAA: std vs. 1 h (R2=0.85) and std vs. 2 h (R2=0.83), (2) ICA512AA: std vs. 1 h (R2=0.85) and std vs. 2 h (R2=0.84), and (3) mIAA: std vs. 1 h (R2=0.70) and std vs. 2 h (R2=0.64). Comparison of assay correlation rates between subject cohorts revealed no significant differences. Compared to their respective standard assays, the 1-h RT GAA assay missed 3.2% and identified an additional 1.3% of samples, the 1-h RT ICA512AA assay had no discordant samples, and the 1-h RT mIAA assay missed 7.1% and identified an additional 5.8% of samples. We analysed a series of 777 stored serum samples from cadaveric donors. Two of 777 (0.25%) were positive for two autoantibodies (both GAA and ICA512AA) and 23 of 777 (3.0%) one autoantibody (11 IAA; 12 GAA). The rapid analysis for all three autoantibodies could be completed in less than 3 h with comparable concordance rates to the more time-consuming standard assays, making these assays an attractive option for organ donor screening to identify potential pancreata for immunopathogenetic research.
本研究的目的是开发并评估针对谷氨酸脱羧酶65自身抗体（GAA）、胰岛细胞抗原512β细胞分化簇/胰岛抗原2（ICA512AA）和胰岛素（微小胰岛素自身抗体，mIAA）的快速检测方法，作为识别有患糖尿病高风险的尸体胰腺供体的潜在工具。该研究纳入了154名新发糖尿病患者、糖尿病前期患者和健康对照者。通过三种独立检测评估受试者的这三种自身抗体：（1）标准（std）检测，在4℃孵育24小时或72小时（分别为联合GAA/ICA512AA或mIAA），（2）在室温（RT）孵育1小时的快速检测，以及（3）在RT孵育2小时的快速检测。还对777名器官供体的血清样本进行了这三种自身抗体的评估，并对标准检测中的所有阳性样本进行了1小时孵育检测。简单线性回归分析显示，对于所有三种自身抗体，标准检测与快速检测之间具有良好的相关性，如下：（1）GAA：标准检测与1小时检测（R2 = 0.85）以及标准检测与2小时检测（R2 = 0.83），（2）ICA512AA：标准检测与1小时检测（R2 = 0.85）以及标准检测与2小时检测（R2 = 0.84），（3）mIAA：标准检测与1小时检测（R2 = 0.70）以及标准检测与2小时检测（R2 = 0.64）。受试者队列之间检测相关性率的比较未发现显著差异。与各自的标准检测相比，1小时RT GAA检测漏检了3.2%的样本，并额外识别出1.3%的样本，1小时RT ICA512AA检测没有不一致的样本，1小时RT mIAA检测漏检了7.1%的样本，并额外识别出5.8%的样本。我们分析了一系列来自尸体供体的777份储存血清样本。777份样本中有2份（0.25%）两种自身抗体（GAA和ICA512AA均为阳性），777份样本中有23份（3.0%）一种自身抗体阳性（11份IAA；12份GAA）。对所有三种自身抗体的快速分析可在不到3小时内完成，与耗时更长的标准检测具有相当的一致性率，这使得这些检测成为器官供体筛查的一个有吸引力的选择，以识别用于免疫发病机制研究的潜在胰腺。
