Abstract
Leptospirosis is a potentially fatal waterborne infection caused by Leptospira interrogans, impacting both humans and animals in tropical regions. However, current diagnostic methods for detecting pathogenic Leptospira have sensitivity, cost, and time limitations. Therefore, there is a critical need for a rapid, sensitive, and cost-effective detection method. This study presents the development of an aptamer-based assay for pathogenic Leptospira detection. Aptamers targeting Leptospira were generated using the SELEX method and screened for binding affinity with major Leptospiral outer membrane proteins through in silico analysis. The aptamer with the highest binding affinity was selected for further evaluation. To enable visual detection, the aptamer was conjugated to gold nanoparticles (AuNPs), resulting in a colorimetric response in the presence of L. interrogans. The aptamer-AuNP-based colorimetric assay exhibited a detection limit of 57 CFU/mL and demonstrated high specificity and reproducibility in detecting pathogenic Leptospira in water samples. This aptamer-based assay represents a significant advancement in leptospirosis diagnostics, offering a rapid, sensitive, and cost-effective approach for detecting pathogenic Leptospira. Its potential for epidemiological applications, such as outbreak source identification and improved prevention, diagnosis, and treatment, particularly in resource-limited settings, highlights its importance in addressing the challenges associated with leptospirosis.