This work proposed the diagnosis of iron deficiency anemia (IDA) and sickle cell disease (SCD) in human blood caused by iron deficiency and hemoglobin S (HbS), which are among the most common anemias, by means of Raman spectroscopy. Whole blood samples from patients diagnosed with IDA and HbS, as well as from normal subjects (HbA), were obtained and submitted to Raman spectroscopy (830 nm, 150 mW, 400–1800 cm−1 spectral range, 4 cm−1 resolution). Difference spectra of IDA–HbA showed spectral features of hemoglobin with less intensity in the IDA, whereas the difference spectra of SCD–HbA showed spectral features of deoxyhemoglobin increased and of oxyhemoglobin decreased in SCD. An exploratory analysis by principal components analysis (PCA) showed that the peaks referred to oxy- and deoxyhemoglobin markedly differentiated SCD and HbA, as well as the increased amount of hemoglobin features in the SCD group, suggesting increased erythropoiesis. The IDA group showed hemoglobin features with lower intensities as well as peaks referred to the iron bonding to the porphyrin ring with reduced intensities when compared to the HbA. Discriminant analysis based on partial least squares (PLS-DA) and PCA (PCA-DA) showed that the IDA and SCD anemias could be discriminated from the HbA spectra with 95.0% and 93.8% of accuracy, for the PLS and PCA respectively, with sensitivity/specificity of 93.8%/95.7% for the PLS-DA model. The iron depletion and the sickling of erythrocytes could be identified by Raman spectroscopy and a spectral model based on PLS accurately discriminated these IDA and SCD samples from the normal HbA.
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L. Silveira Jr. acknowledges FAPESP (São Paulo Research Foundation) for granting the Raman spectrometer (Grant No. 2009/01788-5). L. Silveira Jr. and A. B. Fernandes acknowledge CNPq (National Council for Scientific and Technological Development) for financial support (Grant No. 460014/2014-5). L. Silveira Jr. acknowledges CNPq for the Productivity fellowship (Process No. 306344/2017-3).
Conflict of interest
The authors declare that they have no conflict of interest.
Authors declare compliance with ethical standards when using human blood samples, being the study approved by the Committee for Ethics in Research of Universidade Brasil, Protocol No. 1.578.298 (CAAE No. 53543516.4.0000.5494) following Brazilian regulations for use of human subjects or materials in research (Brazilian Ministry of Health, National Health Council, Resolution CNS No. 466/2012) and have been performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments.
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da Silva, W.R., Silveira, L. & Fernandes, A.B. Diagnosing sickle cell disease and iron deficiency anemia in human blood by Raman spectroscopy. Lasers Med Sci 35, 1065–1074 (2020). https://doi.org/10.1007/s10103-019-02887-1
- Raman spectroscopy
- Sickle cell disease
- Iron deficiency anemia