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用于定量血小板相关IgG检测的血小板对照品的制备。

The production of platelet controls for assays quantitating platelet-associated IgG.

作者信息

Denomme G, Kelton J G

出版信息

Transfusion. 1983 Nov-Dec;23(6):516-8. doi: 10.1046/j.1537-2995.1983.23684074275.x.

DOI:10.1046/j.1537-2995.1983.23684074275.x
PMID:6649029
Abstract

A variety of assays are available for measuring platelet-associated IgG (PAIgG) but the complexity of these assays increases the potential for technical errors. These errors are difficult to detect and, if possible, known positive and negative control platelets should be included with each run. However, patient platelets with elevated levels of IgG are seldom available. This report describes a method for producing positive control platelets that can be labeled with differing amounts of IgG. Normal serum IgG (Cohn fraction II) was incubated with washed 2 percent formalin-fixed platelets for 60 minutes at 37 degrees C. The amount of IgG on the platelets was proportional to the concentration of soluble IgG and the number of incubations. Normal platelet IgG levels were 1.2 +/- 0.1 fg per platelet (mean +/- SEM, n = 34) and positive control platelets had 4.6 +/- 0.2 (n = 12) or 8.4 +/- 0.4 (n = 7). There was no change in the level of PAIgG when stored at 4 degrees C for 1 week, although there was a 28 percent loss in recoverable platelets. When mixed 1:1 with 60 percent glycerol and stored at -70 degrees C, the level of PAIgG was stable for 3 months, with less than 12 percent platelet loss on recovery (n = 12). These positive control platelets have proved useful for monitoring assay performance.

摘要

有多种检测方法可用于测量血小板相关IgG(PAIgG),但这些检测方法的复杂性增加了技术误差的可能性。这些误差很难检测到,如有可能,每次检测都应包含已知的阳性和阴性对照血小板。然而,很少能获得IgG水平升高的患者血小板。本报告描述了一种制备可标记不同量IgG的阳性对照血小板的方法。将正常血清IgG(科恩II组分)与洗涤后的2%福尔马林固定血小板在37℃孵育60分钟。血小板上的IgG量与可溶性IgG的浓度和孵育次数成正比。正常血小板的IgG水平为每血小板1.2±0.1 fg(平均值±标准误,n = 34),阳性对照血小板的IgG水平为4.6±0.2(n = 12)或8.4±0.4(n = 7)。在4℃储存1周时,PAIgG水平没有变化,尽管可回收血小板损失了28%。当与60%甘油按1:1混合并在-70℃储存时,PAIgG水平在3个月内保持稳定,回收时血小板损失少于12%(n = 12)。这些阳性对照血小板已被证明对监测检测性能很有用。

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