A label-free DNAzyme-based colorimetric sensor for the detection of Leptospira interrogans

Abstract

Leptospirosis is a neglected zoonosis caused by a pathogenic spirochete Leptospira. Diagnosis of leptospirosis in the early stage is difficult and can be easily confused with other infections. The existing detection methods are considered chronophagous and labor-intensive. Leptospira survives in the kidney tubules of reservoir animals such as rodents and shed into the environment through their urine. In this study, we developed an Aptamer-DNAzyme-based biosensor for detecting pathogenic Leptospira in environmental water samples. The cell-specific aptamer with an extensive affinity binds to the cell surface proteins to detect the Leptospira interrogans. The DNAzyme that mimics as a peroxidase enzyme, acts as a transducing agent in the colorimetric reaction positively conditioned by the presence of L. interrogans. The Leptospira-specific aptamer coupled with DNAzyme is coated onto carbon nanotubes, to provide a cost-effective nanomaterial-based detection platform. L. interrogans contamination in the samples is detected with a color change of a peroxidase substrate, ABTS. The dissociation constant of the aptazyme was found to be 356.6 nM. The aptazyme system was able to detect up to 119 CFU/mL of L. interrogans exhibiting a high range of selectivity towards the pathogenic spirochete. This simple detection methodology makes the system promising for the environmental monitoring of L. interrogans.

Keywords: Leptospira; Aptasensor; Aptazyme; Carbon nanotubes; DNAzyme; Hemin G-quadruplex.