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I-STOD: a new standardization method for analysing indirect-ELISA results of a schistosomiasis field study

Published online by Cambridge University Press:  11 February 2008

J. LUO
Affiliation:
Department of Mathematics and Computer Science, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
J. XU
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
Y. ZHANG
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
H. SHAN
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
S. ZHANG
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
M. ZHANG
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
X. TU
Affiliation:
Department of Mathematics and Computer Science, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
M. JI
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
F. CHEN
Affiliation:
Department of Epidemiology and Statistics, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
P. M. KNOPF
Affiliation:
Center for International Health Research, Rhode Island Hospital, Providence, RI02903, USA Emeritus Prof. of Medical Science, Department of Molecular Microbiology and Immunology, Brown University, Providence, RI02912, USA
J. KURTIS
Affiliation:
Center for International Health Research, Rhode Island Hospital, Providence, RI02903, USA
G. WU
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
H.-W. WU*
Affiliation:
Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina
*
*Corresponding author: Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu, 210029, PRChina. Tel: +86 25 86862793. Fax: +86 25 86862793. E-mail: haiwei@njmu.edu.cn, Haiwei_Wu@Brown.edu

Summary

Variability among samples analysed using the same ELISA protocol generates ambiguity in deciding which assay best quantifies the protein concentration. In this study, we propose a standardization method, called I-STOD (Improved STandardization method for Optical Density), for the transformation of OD values on different plates into relative concentrations of the antibody levels being assessed. We derived an equation relating OD values of different test samples to antibody levels according to the multi-stage reaction dynamics of the indirect-ELISA. Using serum samples from a Schistosomiasis japonica endemic area, we evaluated the fitness of the I-STOD model to experimental data of a standard reference serum in comparison with 5 other models. Calibration curves fitted by the I-STOD method judged to be superior, based on adjusted R2 (adjusted R2>0·99 on 22 out of 26 plates) values. The CV (coefficient of variation) value of the results between multi-well plates and the number of plates with OD values beyond the control range in Shewhart charts also demonstrate that the I-STOD method is a powerful tool which can greatly improve the comparability of results on different multi-well ELISA plates. We conclude that a standardization method is certainly necessary for antibody levels detected in order to properly illustrate clinical differences.

Type
Original Articles
Copyright
Copyright © Cambridge University Press 2008

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References

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