STATISTICAL CONSIDERATIONS IN THE QUANTITATION OF SERUM IMMUNOGLOBULIN LEVELS USING THE ENZYME-LINKED-IMMUNOSORBENT-ASSAY (ELISA)
STATISTICAL CONSIDERATIONS IN THE QUANTITATION OF SERUM IMMUNOGLOBULIN LEVELS USING THE ENZYME-LINKED-IMMUNOSORBENT-ASSAY (ELISA)
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DOI：
10.1016/0022-1759(87)90289-4
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发表时间：
1987-11-05
影响因子：
2.2
通讯作者：
TRYPHONAS, H
中科院分区：
文献类型：
--
作者：
KARPINSKI, KF;HAYWARD, S;TRYPHONAS, H
在微夹心技术中，用涂有适当稀释的人免疫球蛋白和免疫球蛋白M的抗血清的微孔板，将恒河猴(Macaca Mulatta)参考血清与过氧化物酶标记的抗人免疫球蛋白和免疫球蛋白M的单特异性抗血清的连续两倍稀释度反应，评价了几种分析酶联免疫吸附试验数据的方法。将光密度表示为对数血清稀释度的线性函数被证明是不适当的，并且可能具有误导性。4参数LOGIT的加权非线性最小二乘分析被证明是不合适的，因为它需要指定某种任意的方差函数，并且权重估计值随所用函数的不同而有很大差异。将光密度表示为四参数Logistic函数，并在对数尺度上进行估计，是用ELISA法测定抗原或抗体浓度的最合适的方法。
Several methods for analyzing ELISA data have been evaluated using optical density values derived by reacting serial two-fold dilutions of a rhesus monkey (Macaca mulatta) reference serum with dilutions of peroxidase-conjugated monospecific antisera to human IgG and IgM in the micro-ELISA ''sandwich'' technique using microplates coated with appropriately diluted antisera to human IgG and IgM. Representation of optical density as a linear function of log serum dilution was shown to be inappropriate and potentially misleading. Weighted non-linear least squares analysis of a 4-parameter logit was demonstrated to be inappropriate because it required the specification of a somewhat arbitrary variance function and weight estimates varied substantially depending on the function used. Representation of optical density as a 4-parameter logistic function with estimation carried out on the log scale was shown to be the most appropriate procedure for determining the concentration of antigens or antibodies by the ELISA method.