Department of Genetic Engineering, Faculty of Engineering and Technology, School of Bioengineering, SRM Institute of Science and Technology, SRM Nagar, Kattankulathur, Kanchipuram, Chennai, Tamil Nadu, 603203, India.
Sci Rep. 2023 Aug 17;13(1):13379. doi: 10.1038/s41598-023-40120-w.
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.
钩端螺旋体病是一种由问号钩端螺旋体引起的潜在致命性水传播感染，影响热带地区的人类和动物。然而，目前用于检测致病性钩端螺旋体的诊断方法存在灵敏度、成本和时间限制。因此，迫切需要一种快速、敏感和具有成本效益的检测方法。本研究提出了一种基于适配体的致病性钩端螺旋体检测方法。使用 SELEX 方法生成针对钩端螺旋体的适配体，并通过计算机分析筛选与主要钩端螺旋体外膜蛋白具有高结合亲和力的适配体。选择具有最高结合亲和力的适配体进行进一步评估。为了实现可视化检测，将适配体与金纳米粒子（AuNPs）偶联，在存在 L. interrogans 时会产生比色响应。基于适配体-AuNP 的比色测定法的检测限为 57 CFU/mL，并且在检测水样中的致病性钩端螺旋体时表现出高特异性和重现性。这种基于适配体的检测方法代表了钩端螺旋体病诊断的重大进展，提供了一种快速、敏感和具有成本效益的方法来检测致病性钩端螺旋体。它在流行病学应用中的潜力，如暴发源识别和改进的预防、诊断和治疗，特别是在资源有限的环境中，突出了它在应对与钩端螺旋体病相关的挑战方面的重要性。
