FTIR/ATR Spectroscopy Applied In Wavelength Models Study For Analysis Of Thalassemia Screening Indicators

Thalassemia is a serious hereditary hemolytic disease, and it affects individuals from many parts of the world, including South China. Thalassemia carriers screening is fundamental for preventing and controlling severe thalassemia. Hemoglobin(Hb), mean corpuscular Hb(MCH), and mean corpuscular volume(MCV) are the most important thalassemia screening indicators. However, ancillary reagents and professional operators are always required in conventional detection methods, and are not suitable for screening large populations. In the present study, a rapid quantitative analysis method for reagent-free determination for Hb, MCH and MCV of the human peripheral blood samples was developed with Fourier transform infrared(FTIR) spectrometers and attenuated total reflection(ATR) combined with chemometrics.Based on several divisions of calibration and prediction sets, a stability-dependent frame of calibration, prediction, and validation was conducted. On this basis, moving window partial least squares(MW-PLS) method was improved, and to establish quantitative models and wavelength models for Hb, MCH and MCV analysis in FTIR/ATR, respectively; The optimal MW-PLS wavebands were 1,722–1,504 cm-1 for Hb, 1,653–901 cm-1 for MCH, and 1,562–964 cm-1 for MCV, respectively. Considering the restrictions of position and number of wavelengths caused by costs and material properties in the instrument design, local optimal models that based on the position and number of wavelengths and model sets that equivalent to the optimal MW-PLS models were proposed. We found the public waveband of Hb equivalent model set(1,717–1,510 cm-1) and the equivalent public wavebands of MCH and MCV(1,562–901 cm-1), had equivalent prediction effect with the optimal MW-PLS wavebands, and the model complexity was reduced. Based on optimal MW-PLS models, the prediction root-mean-square error in the validation set(V_SEP) for Hb, MCH and MCV were 7.7 g.L-1、2.28 pg、5.14 f L, respectively, the corresponding sensitivity and specificity for thalassemia achieved 100.0%、96.9%, respectively; Based on equivalent public waveband of equivalent model sets, the V_SEP for Hb, MCH and MCV were 7.6 g.L-1、2.22 pg、5.25 f L, respectively, the corresponding sensitivity and specificity for thalassemia achieved 100.0%、95.3%, respectively.The results confirmed the feasibility of quantitative analysis of Hb, MCH, and MCV using FTIR/ATR spectroscopy combined with MW-PLS method. The proposed model sets could provide valuable references for designing special spectrum analysis instruments. This method could reagent-free determination for Hb, MCH and MCV of the human peripheral blood, and suitable for screening thalassemia in large populations.