Harrison P, Ault K A, Chapman S, Charie L, Davis B, Fujimoto K, Houwen B, Kunicka J, Lacombe F, Machin S, Raynor R, van Hove L, van Assendelft O W
University College London, London, England.
Am J Clin Pathol. 2001 Mar;115(3):448-59. doi: 10.1309/91PR-E4G6-XBAF-N8DY.
A multinational interlaboratory task force explored the important variables of platelet reference counting and developed a candidate flow cytometric reference method based on the RBC/platelet ratio. A multicenter comparison was performed to determine whether the method met the necessary criteria and was precise enough to be recommended as a new reference method. Each laboratory analyzed serial dilutions of normal specimens, stabilized material, and at least 60 patient specimens with a range of platelet counts from 1 to 400 x 10(3)/microL (1-400 x 10(9)/L). Pooled analysis of the serial dilutions showed that RBC-platelet and RBC-RBC coincidence events became negligible at sufficiently high dilutions (i.e., > 1:1,000). All laboratories demonstrated excellent intra-assay and acceptable interlaboratory precision. Two antibodies (CD61 and CD41) were used for identifying platelets and individually gave acceptable results, but in a minority of samples, staining differences were observed. The optimum method thus uses a double-labeling procedure with a final dilution factor of 1:1,000. The study demonstrated that this method meets the criteria for a reference platelet count.
一个跨国实验室间特别工作组探讨了血小板参考计数的重要变量，并基于红细胞/血小板比率开发了一种流式细胞术候选参考方法。进行了多中心比较，以确定该方法是否符合必要标准，以及是否精确到足以被推荐为一种新的参考方法。每个实验室分析了正常标本、稳定材料的系列稀释液，以及至少60份血小板计数范围为1至400×10³/微升（1-400×10⁹/升）的患者标本。系列稀释液的汇总分析表明，在足够高的稀释度（即>1:1000）下，红细胞-血小板和红细胞-红细胞重合事件可忽略不计。所有实验室均表现出出色的批内精密度和可接受的实验室间精密度。使用两种抗体（CD61和CD41）识别血小板，各自给出了可接受的结果，但在少数样本中观察到染色差异。因此，最佳方法是采用最终稀释因子为1:1000的双标记程序。该研究表明，此方法符合血小板参考计数的标准。
