EAACI task force report: A consensus protocol for the basophil activation test for collaboration and external quality assurance.
EAACI task force report: A consensus protocol for the basophil activation test for collaboration and external quality assurance.
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EAACI 工作组报告：用于协作和外部质量保证的嗜碱性粒细胞激活测试的共识协议。
作者：
Pascal,M;Edelman,SM;Nopp,A;Möbs,C;Geilenkeuser,WJ;Knol,EF;Ebo,DG;Mertens,C;Shamji,MH;Santos,AF;Patil,S;Eberlein,B;Mayorga,C;Hoffmann,HJ
| 291 PASCAL等. performance. 1，2法规批准和临床实施需要实验室方案、细胞仪设置和结果解释的广泛标准化。3欧洲国家当局要求由独立于测试供应商的机构对现代诊断实验室的性能进行外部质量保证（EQA），以满足ISO 15189：2012、15189：2013和9001：2015。基于2017年对59个进行BAT的欧洲实验室进行的在线调查4，5（在线补充;在线调查结果），2018年成立了一个工作组（TF），为BAT-EQA奠定基础。组织Round Robins（RR），从德国波恩（Bonn，DE）向7-10个欧洲中心运送7次，每次运送2名供体，并提供隔夜快递服务。将每份样本分成两等份：（i）为尽量减少变异，在运输前，用1 μL抗Fc εRI抗体/mL血液活化匿名供体的血液嗜碱性粒细胞，并用0.2 mL Transfix（Cytomark，UK）/mL血液稳定，以稳定活化的嗜碱性粒细胞长达24 h，用于染色6;（ii）在参与的实验室研究中心纳入新鲜血液进行刺激和染色。TF成员在第三次运输后开会，就BAT方案达成共识（在线补充;内部BAT的拟议SOP）。数据分析从散点图中相关区域的鉴定开始（图1A），然后按照不同策略鉴定具有相关标志物的嗜碱性粒细胞，例如，使用低SSC和仅CD 193或CD 193/CD 123或CD 193/CD 203 c（图1B）。
| 291 PASCAL et al. performance. 1, 2 Regulatory approval and clinical implementation require extensive standardization of laboratory protocols, cytometer settings, and results interpretation. 3 European national authorities require External Quality Assurance (EQA) of the performance of modern diagnostic laboratories by agencies independent of test suppliers to meet ISO 15189: 2012, 15189: 2013, and 9001: 2015. Based on an online survey among 59 responding European laboratories performing BAT in 2017 4, 5 (Online Supplement; Results of the online survey), a Task Force (TF) was launched in 2018 to create the basis for a BAT-EQA. Round Robins (RR) were organized with seven shipments of two donors each to 7–10 European centers with overnight courier service from Bonn, DE. Each sample was split into two aliquots:(i) To minimize variation, prior to shipment, blood basophils of anonymous donors were activated with 1 μL anti-FcεRI antibody/mL of blood and stabilized with 0.2 mL Transfix (Cytomark, UK) per mL of blood to stabilize activated basophils up to 24 h for staining 6;(ii) Fresh blood was included for stimulation and staining at the participating laboratory sites.TF members met after the third shipment to reach consensus on a protocol for BAT (Online Supplement; Proposed SOP for in house BAT). Data analysis started with identification of the relevant region in a scatter plot (Figure 1A), followed by identification of basophils with the relevant markers following different strategies, for instance, using low SSC and CD193 only or CD193/CD123 or CD193/CD203c (Figure 1B).