Development of An Apta Biosensor for Rapid Detection of Leptospira Interrogans

Abstract

Aptamers, single-stranded DNA or RNA molecules, are gaining recognition newlinein biomedical applications due to their ability to bind to specific targets with high affinity and specificity, along with their rapid analytical capabilities. However, existing newlinediagnostic methods for detecting pathogenic Leptospira, the causative agent of leptospirosis, suffer from significant limitations, including low sensitivity, lack of specificity, and high costs, particularly in the early stages of infection. This study addresses these challenges by leveraging the unique binding properties of aptamers to develop a more effective diagnostic tool. To bridge the gaps in current Leptospira detection methods, a SELEX-based newlineapproach was employed to generate and identify aptamers with high binding affinity for pathogenic Leptospira. Following the selection process, bioinformatic analyses, newlineincluding sequencing, secondary structure prediction, and docking studies, were conducted to identify the most promising aptamers. Among the identified candidates, newlineaptamer LAP1 demonstrated the highest structural stability and binding affinity toward newlineLeptospiral outer membrane protein, as confirmed by in silico docking studies. This was further validated by labeling LAP1 with a fluorophore, Fluorescein isothiocyanate (FITC), to assess its binding performance in vitro newline