title: Validation of a method for predicting the precision, limit of detection and range of quantitation in competitive ELISA.
abstract: The mathematical model for predicting the precision, limit of detection (LOD) and range of quantitation (ROQ) in a competitive enzyme-linked immunosorbent assay (ELISA) proposed by Hayashi et al. (Anal. Chem., 2004, 76, 1295) was validated. The model describes the relative standard deviation (RSD) of concentration estimates by the RSDs of pipetting volumes of analyte, enzyme-conjugated antigen, antibody and substrate solutions, and the standard deviation (SD) of inherent absorbances between the wells in an ELISA plate. For 6 kinds of direct competitive ELISA kits, the LOD and ROQ predicted by the model agreed well with those obtained by experiments with real samples. It was also confirmed that the model is applicable to the prediction of uncertainty that depends on the pipetting error of the viscous antiserum solution. The model was demonstrated to be useful for estimating the LOD and ROQ of competitive ELISA.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
authors.forename: Dong Hwan
authors.initials: DH
authors.lastname: Choi
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Katakura
authors.forename: Rieko
authors.initials: R
authors.lastname: Matsuda
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Masato
authors.initials: M
authors.lastname: Hirobe
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Goda
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Ninomiya
authors.forename: Suteaki
authors.initials: S
authors.lastname: Shioya
mesh_terms: D001577:Benzophenones / Q000032:analysis; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D004958:Estradiol / Q000032:analysis; D004967:Estrogens / Q000032:analysis; D004970:Estrone / Q000032:analysis; D005278:Fenitrothion; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D013677:Medical Laboratory Science / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D001577:Benzophenones; D004967:Estrogens; D007106:Immune Sera; D004970:Estrone; D004958:Estradiol; C482525:2,2'-dihydroxy-4,4'-dimethoxybenzophenone-5,5'-disulfonic acid; D005278:Fenitrothion