Innovative robust basophil activation test using a novel gating strategy reliably diagnosing allergy with full automation. Issue 12 (28th May 2021)

• Record Type: Journal Article
• Authors: Behrends, Jochen; Schwager, Christian; Hein, Martina; Scholzen, Thomas; Kull, Skadi; Jappe, Uta
• Is Part Of: Allergy. Volume 76:Issue 12(2021)
• Page Start: 3776
• Page End: 3788
• Publication Date: 2021-05-28
• Subjects: allergy diagnostic -- clinical trials -- oleosins -- peanut allergy -- sensitization; Allergy -- Periodicals; 616.97
• DOI: 10.1111/all.14900
• Languages: English
• ISSNs: 0105-4538

Abstract

Abstract: 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 (FcεRIα, 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 (FcεRIα, 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°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°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. Abstract : BAT has limits to be a routine application. Therefore, we developed a robust innovative gating strategy and demonstrated a prolonged time frame for measurements (nationwide ring trial). The enormous laboratory work could significantly be reduced via the automatic sample processing and measurement protocols (automated analysis template using R Studio) to reduce the hands‐on time. Abbreviation: BAT, basophil activation test.