Hemoglobinopathies are among the most common inherited diseases worldwide, affecting approximately 7% of the global population. Despite advances in the standardization and harmonization of methods for HbA1c determination, an increasing number of hemoglobinopathies cause false HbA1c results. One of the common techniques for screening hemoglobinopathies is through high-performance liquid chromatography (HPLC) separation, followed by UV–VIS detection. Although UV–VIS can quantify the hemoglobin fractions, it is unable to identify them. In this study, we use Raman spectroscopy to study the fingerprint spectra of hemoglobin fractions based on which the fractions can be identified. To evaluate the potential of Raman spectroscopy in identifying these fractions, we utilize a range of commercially available hemoglobin fractions, including fetal hemoglobin. We automate the classification process with machine learning approaches such as support vector machines (SVM), fully connected neural networks (NN), k-Nearest Neighbors (KNN), Decision Trees (DT), and Bernoulli Naive Bayes (BNB). These models are fine-tuned and optimized to classify the hemoglobin fractions and achieve test accuracies of 98.2% and 98.5%, respectively. Our research highlights the potential of Raman spectroscopy as an identification tool when combined with HPLC.
Hemoglobinopathies are the most common genetic disorders caused by a mutation in the genes encoding for one of the globin chains and leading to structural (hemoglobin [Hb] variants) or quantitative defects (thalassemias) in hemoglobin. Early diagnosis and characterization of hemoglobinopathies are essential to avoid severe hematological consequences in the offspring of healthy carriers of a mutation. Despite being extensively studied, hemoglobinopathies continue to provide a diagnostic challenge. Sickle-cell hemoglobin (HbS) is the most common and clinically significant hemoglobin variant among all Hb variants. To overcome the challenge of diagnosing Hb variants, we propose the use of Surface-Enhanced Raman Spectroscopy (SERS). SERS is a powerful label-free tool for providing fingerprint structural information of analyses. It can rapidly generate the spectral signature of samples. This study investigates the structural differences between HbS and normal Hb using gold nanopillar SERS substrates with a leaning effect. The SERS spectra of Hb variants showed subtle spectral differences between HbS and normal Hb located in the valine (975 cm-1) and glutamic acid (1547 cm-1) band, reflecting the amino acid substitution in the HbS β-globin chain. We also automated the identification of HbS and normal Hb with principal component analysis (PCA) combined with support vector machine (SVM) and linear discriminant analysis (LDA) classifiers, leading to an accuracy of 98% and 96%, respectively. This study demonstrated that SERS can provide a fast, highly sensitive, noninvasive, and accurate detection module for the diagnosis of Sickle-cell disease and potentially other hemoglobinopathies.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.