期刊
ALLERGY
卷 76, 期 12, 页码 3776-3788出版社
WILEY
DOI: 10.1111/all.14900
关键词
allergy diagnostic; clinical trials; oleosins; peanut allergy; sensitization
资金
- German Research Foundation (DFG) [JA 1007/2-1, JA 1007/2-3]
- Federal Ministry of Education and Science (German Center for Lung Research (DZL))
This study successfully established a highly specific, high-throughput, and automated basophil activation test (BAT) through a novel gating strategy, optimized storage conditions, and automated measurement, demonstrating efficient detection of allergies.
Background Allergy is one of the most common chronic diseases in Europe. Therefore, an increased need for specific and sensitive diagnostic tests that truly detect allergy exists. This study aimed at establishing a highly specific high-throughput and automated basophil activation test (BAT) that proves the existence of an allergy with utmost probability. Methods BAT from 1104 samples was analyzed; a novel gating strategy with three antibodies (Fc epsilon RI alpha, CD203c, CD63) was established and compared with our published protocol (12 antibodies). Based on the novel gating strategy, storage conditions, automated measurement, and analyses using R (1376 samples out of 1389) were optimized to set up a high-throughput BAT. Results No differences in sensitivity and specificity were found between the novel three antibody (Fc epsilon RI alpha, CD203c, CD63) and the 12 antibody gating strategy or between automated and manually analyzed samples (saving up to 90% of labor time). The time frame for basophil activation measurement after blood donation has been extended considerably (whole blood storage <= 7 days (RT) and 17 days (4 degrees C) prior to BAT preparation and measurement). Respective storage conditions were optimized for samples after stimulation, staining, and preparation (<= 7 days (RT) and 28 days (4 degrees C)). These achievements were confirmed by a nationwide ring trial showing robustness and applicability of our BAT on a variety of flow cytometers. Conclusion Our considerable optimizations overcame the hurdles that until now prevented the BAT from being used as high-throughput allergy diagnostic test in routine laboratories and shall allow for collaborative studies between clinics and research centers.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据