Validation of Urinary Metabolites as Diagnostic Biomarkers for Dengue Infection

Abstract

Dengue, a mosquito-borne viral infection, presents significant challenges to global health, particularly in tropical and subtropical regions. Early and accurate diagnosis is essential for effective patient management. Urine-based metabolic profiling has emerged as a promising method for identifying disease biomarkers. In a preliminary study (n=10), we identified nine urinary metabolites associated with dengue infection namely heptacosane, hexadecane, 2-bromooctane, tetradecane, pentadecane, 2,9-dimethyldecane, 2,4-bis (1,1-dimethylethyl) phenol, hexyl octyl ester sulphuric acid and 2-benzoyl methyl ester benzoic acid, using Gas Chromatography Mass Spectrometry (GC-MS). To identify the most sensitive biomarkers in dengue infection, this study aims to assess the reproducibility of the targeted urinary metabolites in dengue infection using GC-MS. We compared the presence of these nine metabolites in 30 urine samples of both dengue-infected patients and 30 controls using GC-MS based on the retention time (RT) and similarity index (SI). We also quantified the levels of selected metabolites based on the peak area. We found that two out of the nine targeted metabolites, specifically hexadecane and pentadecane, were present in the urine of dengue-infected patients compared to the healthy controls (p<0.01 and p<0.01 respectively). Hexadecane showed a RT of 20.95 ± 2.23 min, SI of 85.50 ± 5.00 % and peak area of 1360566.25 ± 1066618.37 a.u, while pentadecane had a RT of 24.07 ± 3.35 min, SI of 86.00 ± 4.55 % and peak area of 853458.25 ± 523318.12 a.u. These results highlight the potential of GC-MS as a reliable and reproducible detection tool for urinary metabolites dengue infection. The identification of key metabolites enhances our understanding of the metabolic changes associated with dengue infection. It also paves the way for the development of rapid, non-invasive urine-based dengue diagnostic tests. Such tests are more patient-friendly, self-administrable and widely accessible, thereby facilitating more timely diagnosis and optimum patient care.