Rodbard D, Lenox R H, Wray H L, Ramseth D
Clin Chem. 1976 Mar;22(3):350-8.
We have developed practical methods for evaluating the magnitude of the random errors in radioimmunoassay dose--response variables, and the relationship between this error and position on the dose--response curve. This is important: to obtain appropriate weights for each point on the dose--response curve when utilizing least-squares curve-fitting methods; to evaluate whether the standards and the unknowns are subject to error of the same magnitude; for quality-control purposes; and to study the sources of errors in radioimmunoassay. Both standards and unknowns in radioimmunoassays for cAMP and cGMP were analyzed in triplicate. The same mean (Y), sample standard deviation, sy, and variance (2-y) of the response variable were calculated for each dose level. The relationship between s 2-y and y was calculated utilizing several models. Results for standards and unknowns from several assays were pooled, and a curve smoothing procedure was used to minimize random sampling errors. This pooling increased the reliability of the analysis, and confirmed the presence of the theoretically predicted nonuniformity of variance. Thus, the calculation of results from these radioimmunoassays should utilize a weighted least-squares curve-fitting program. These analyses have been computerized, and can be used as a "pre-processor" for programs for routine analysis of results of radioimmunoassay.
我们已经开发出实用方法，用于评估放射免疫分析剂量-反应变量中随机误差的大小，以及这种误差与剂量-反应曲线上位置的关系。这一点很重要：在使用最小二乘曲线拟合方法时，为剂量-反应曲线上的每个点获得合适的权重；评估标准品和未知样品是否受到相同大小误差的影响；用于质量控制目的；以及研究放射免疫分析中的误差来源。对环磷酸腺苷（cAMP）和环磷酸鸟苷（cGMP）放射免疫分析中的标准品和未知样品均进行了三次重复分析。针对每个剂量水平计算了反应变量的相同均值（Y）、样本标准差（sy）和方差（s²y）。利用几种模型计算了s²y与y之间的关系。汇总了几次分析中标准品和未知样品的结果，并使用曲线平滑程序将随机抽样误差降至最低。这种汇总提高了分析的可靠性，并证实了理论预测的方差不均匀性的存在。因此，这些放射免疫分析结果的计算应使用加权最小二乘曲线拟合程序。这些分析已经计算机化，可作为放射免疫分析结果常规分析程序的“预处理器”。
