Structural characterization of DNA amplicons by ATR-FTIR spectroscopy as a guide for screening metainflammatory disorders in blood plasma

• Published in: Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy Vol. 310; p. 123897
• Main Authors: Mateus Pereira de Souza, Nikolas, Kimberli Abeg da Rosa, Dhuli, de Moraes, Caroline, Caeran, Mariana, Bordin Hoffmann, Mairim, Pozzobon Aita, Eduardo, Prochnow, Laura, Lya Assmann da Motta, Anna, Antonio Corbellini, Valeriano, Rieger, Alexandre
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 05.04.2024
• Subjects: Ataxia Telangiectasia Mutated Proteins; ATR-FTIR spectroscopy; Cell-free DNA; Discriminant Analysis; Humans; Metainflammation; Phosphates; Plasma; Spectroscopy, Fourier Transform Infrared - methods; Sugars; Variable selection; Cell-free DNA; ATR-FTIR spectroscopy; Metainflammation; Variable selection
• ISSN: 1386-1425; 1873-3557
• DOI: 10.1016/j.saa.2024.123897

[Display omitted] •100% accuracy for MetS vs. control in blood plasma guided by IR peaks of DNA.•Curve fitting in the DNA IR range 1475–1188 cm−1 with adjusted-R2 of 0.993.•97 % accuracy for MetS vs. control at 1475–1188 cm−1 by PCA with discriminant analysis. Attenuated total reflectance (ATR) Fourier transform infrared (FTIR) spectroscopy is a promising rapid, reagent-free, and low-cost technique considered for clinical translation. It allows to characterize biofluids proteome, lipidome, and metabolome at once. Metainflammatory disorders share a constellation of chronic systemic inflammation, oxidative stress, aberrant adipogenesis, and hypoxia, that significantly increased cardiovascular and cancer risk. As a result, these patients have elevated concentration of cfDNA in the bloodstream. Considering this, DNA amplicons were analyzed by ATR-FTIR at 3 concentrations with 1:100 dilution: (IU/mL): 718, 7.18, and 0.0718. The generated IR spectrum was used as a guide for variable selection. The main peaks in the biofingerprint (1800–900 cm−1) give important information about the base, base-sugar, phosphate, and sugar-phosphate transitions of DNA. To validate our method of selecting variables in blood plasma, 38 control subjects and 12 with metabolic syndrome were used. Using the wavenumbers of the peaks in the biofingerprint of the DNA amplicons, was generated a discriminant analysis model with Mahalanobis distance in blood plasma, and 100 % discrimination accuracy was obtained. In addition, the interval 1475–1188 cm−1 showed the greatest sensitivity to variation in the concentration of DNA amplicons, so curve fitting with Gaussian funcion was performed, obtaining adjusted-R2 of 0.993. PCA with Mahalanobis distance in the interval 1475–1188 cm−1 obtained an accuracy of 96 % and PLS-DA modeling in the interval 1475–1088 cm−1 obtained AUC = 0.991 with sensitivity of 95 % and specificity of 100 %. Therefore, ATR-FTIR spectroscopy with variable selection guided by DNA IR peaks is a promising and efficient method to be applied in metainflammatory disorders.