COMPARISONS OF STANDARD CURVE-FITTING METHODS TO QUANTITATE NEISSERIA-MENINGITIDIS GROUP-A POLYSACCHARIDE ANTIBODY-LEVELS BY ENZYME-LINKED-IMMUNOSORBENT-ASSAY
COMPARISONS OF STANDARD CURVE-FITTING METHODS TO QUANTITATE NEISSERIA-MENINGITIDIS GROUP-A POLYSACCHARIDE ANTIBODY-LEVELS BY ENZYME-LINKED-IMMUNOSORBENT-ASSAY
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DOI：
10.1128/jcm.29.7.1439-1446.1991
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发表时间：
1991-07-01
影响因子：
9.4
通讯作者：
CARLONE, GM
中科院分区：
文献类型：
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作者：
PLIKAYTIS, BD;TURNER, SH;CARLONE, GM
我们研究了几种更常用的模型(LOG-LOG，两种形式的LOGIT-LOG，以及四参数Logistic-LOG变换)，以使用标准化的酶联免疫吸附试验(ELISA)来形成标准或校准曲线。测量并比较每种功能的测定范围、准确度和误差。通过计算已知浓度的系列稀释标准参考血清的抗体浓度来估计对A组脑膜炎奈瑟氏菌多糖的抗体水平。每个函数都达到了很高的平方相关系数(R2>；0.97)，表明在形成标准曲线时具有很高的准确性。然而，当将预测的抗体浓度与已知值进行比较时，对数-对数函数表现出的精度最低，在几个稀释度上出现了极端的误差百分比。在较小的标准稀释范围内，部分指定的LOGIT-LOG变换比LOG-LOG模型执行得更好。这表明，高R2本身并不是标准曲线准确性的可靠衡量标准。在调查的方法中，Logistic-log和完全指定的Logit-log函数是形成标准曲线和从标准曲线内插抗体浓度最准确的模型。完全指定的Logit-log函数的精度在很大程度上取决于两个未知量的精确指定，即在将模型与一组典型的校准数据进行匹配之前，对两个未知量的精确指定，即在零浓度和无限浓度下的光学密度。四参数Logistic-log函数是检测A群脑膜炎奈瑟菌总多糖抗体的首选方法。该函数在估计标准曲线之前不需要预先指定任何参数，并且这四个参数很容易用可识别的物理量来解释。该模型还具有最易于可视化的优点，因为它不包含光学密度标度的复杂变换。
We examined several of the more commonly used models (log-log, two forms of the logit-log, and the four-parameter logistic-log transformations) for forming standard or calibration curves by using a standardized enzyme-linked immunosorbent assay (ELISA). Assay range, accuracy, and error for each function were measured and compared. Antibody levels to Neisseria meningitidis group A polysaccharide were estimated by calculating antibody concentrations of a serially diluted standard reference serum of known concentration. Each function achieved a high squared correlation coefficient (r2 > 0.97), indicating a high degree of accuracy in forming the standard curves. However, when predicted antibody concentrations were compared with the known values, the log-log function exhibited the least precision, with extreme precentages of error occurring at several dilutions. A partially specified logit-log transformation performed better than the log-log model over a reduced range of standard dilutions. This indicated that a high r2 alone was not a reliable measure of the accuracy of the standard curve. Of the methods surveyed, the logistic-log and fully specified logit-log functions were the most accurate models for forming standard curves and for interpolating antibody concentrations from the standard curve. The accuracy of the fully specified logit-log function is highly dependent on the precise specification of two unknown quantities, the optical densities at zero and infinite concentrations, prior to fitting the model to a typical set of calibration data. The four-parameter logistic-log function was the preferred choice for quantitating N. meningitidis group A total polysaccharide antibody by using a standardized ELISA. The function does not require prespecification of any parameters before estimating the standard curve, and the four parameters are readily interpretable in terms of identifiable physical quantities. This model also has the advantage that it is easiest to visualize since it does not incorporate complex transformations of the optical density scale.