Validation for quantification of immunoglobulins by Fourier transform infrared spectrometry.

摘要

BACKGROUND: The objective of this study was to develop a robust quantification method for simultaneously analyzing molecules in human plasma using the Fourier transform infrared (FT-IR) system with a partial least square (PLS) regression. METHODS: Plasma spectra were analyzed from 4000 to 500 cm(-1) (with 2.0 cm(-1) of resolution and 32 scans), and the molecule concentrations (IgA, IgG, IgM) were measured blindly by using a cross-validation model prepared by PLS analysis of data from 135 samples. RESULTS: There was a significant correlation between the FT-IR predicted concentration and the concentration obtained with the clinical reference method: R(2)=0.98 (IgA), R(2)=0.98 (IgG), and R(2)=0.97 (IgM). The root mean square error of prediction (RMSEP) was 0.05 g.L(-1) (IgA), 0.4 g.L(-1) (IgG), and 0.03 g.L(-1) (IgM). Variability of inter-experimenter reproducibility was less than 2%. The interchangeability of the two methods was studied by using the Bland-Altman method. CONCLUSIONS: Together with PLS analysis, FT-IR spectrometry appears to be an easy-to-use and accurate method to determine multianalyte concentrations in dried human plasma. It could be an alternative tool for rapidly quantifying many molecules after developing a specific predictive model.