Reducing the variation in performance of antibody titrations
Reducing the variation in performance of antibody titrations
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DOI：
10.1043/1543-2165(2008)132
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发表时间：
2008-07-01
影响因子：
4.6
通讯作者：
Dumont, L. J.
中科院分区：
文献类型：
--
作者：
AuBuchon, J. P.;de Wildt-Eggen, J.;Dumont, L. J.
背景。-抗体滴定很难标准化。我们在一项国际研究中调查了详细、统一的抗体滴定程序是否会减少试管和凝胶卡滴度的差异。方法-实验室(n=35)分别按照(I)他们的常规方法；(Ii)详细的、统一的方法；以及(Iii)统一的方法，将血清样本滴定到特定表型的红细胞(D+C-c+E+e-用于抗-D；A(1)用于抗-A)，而不是熟练测试试剂盒中提供的相同表型的红细胞。用1+和w+终点报告了统一的方法结果。使用F检验进行成对的方差统计分析。结果：使用1+终点的统一方法，实验室间的方差没有显著降低。然而，当19个实验室使用w+终点重新分析他们的结果时，在37℃孵育和转换到抗球蛋白(AHG)相后，通过基于试管的统一技术，抗-D和抗-A滴度的差异在统计上显著降低。很少有实验室报告凝胶卡测试中出现w+终点的结果，无法进行分析。由参与实验室提供的针对特定表型的红细胞的滴定似乎没有引入额外的差异。总体而言，基于凝胶卡技术在AHG阶段(1+终点)报告的结果显示，与基于试管的技术相比，差异更小。结论：一种详细、统一的抗体滴定方法，在37℃下，在AHG阶段以试管为基础的方法读取，w+终点降低了实验室间的变异性。
Background.-Antibody titration is difficult to standardize. We investigated whether a detailed, uniform procedure for antibody titration would reduce variation in both tube-based and gel card titres in an international study.Methods.-Laboratories (n = 35) tested proficiency testing material provided by the College of American Pathologists each according to (i) their routine method; (ii) a detailed, uniform method; and (iii) the uniform method titrating the serum sample against a red cell of specified phenotype (D+ C- c+ E+ e- for anti-D; A(1) for anti-A) instead of the red cell of the same phenotype provided in the proficiency testing kit. Uniform method results were reported with 1+ and w+ end-points. Paired statistical analyses of variance were conducted using the F-test.Results.-The variance between laboratories was not significantly reduced with the uniform method using a 1+ end-point. However, a statistically significant reduction in the variance of anti-D and anti-A titres by the tube-based uniform technique after 37 degrees C incubation and conversion to the antiglobulin (AHG) phase was seen when 19 laboratories reanalysed their results using a w+ end-point. Too few laboratories reported results with a w+ end-point in gel card testing to allow analysis. Titration against red cells of the specified phenotype provided by the participating laboratory did not appear to introduce additional variance. Overall, results reported based on the gel card technique at the AHG phase (1+ end-point) showed reduced variance compared to tube-based techniques.Conclusions.-A detailed, uniform method for antibody titration at 37 degrees C and read at the AHG phase in a tube-based method with a w+ end-point reduced interlaboratory variability.