Pathogenic Leptospira Detection in Environmental Contaminant Water Sources by Highly Performance Antibody Absorption Polystyrene Agglutinating Particles

Abstract

Leptospirosis is a re-emerging bacterial zoonotic disease that affects both humans and animals, with a significantly higher incidence in tropical and sub-tropical regions. Disease control, epidemiology, and surveillance rely on a One Health approach, as accurate detection can be applied to humans, animals, and the environment. This study represents the first attempt to develop a method for detecting the pathogenic Leptospira santarosai serovar Shermani based on the latex agglutination reaction. The serological activity of the antibody is examined to achieve a high titer of antibody before adsorption onto polystyrene particles. Using a pH medium of 6.8–7.8, total antibody adsorption of up to 3 mg m−2 is achieved. Particle agglutination is observed after incubating the antibody-adsorbed PS with leptospiral culture for 4 min, revealing a detection limit of 1.7 × 102 leptospires mL−1. Interestingly, the detection limit increased by 1000 times when observing agglutination using spectrophotometer. The test exhibits high specificity with Shermani and shows negligible cross-agglutination with non-pathogenic Leptospira and water-borne bacteria. Agglutination testing in collected water samples from natural sources demonstrates a good correlation with culture technique. This simple and rapid leptospires agglutination detection method can be applied as a screening test in environmental, human, and animal specimens.