Visual and Rapid Detection of Leptospira interrogans Using Multiple Cross-Displacement Amplification Coupled with Nanoparticle-Based Lateral Flow Biosensor.

Abstract

Leptospirosis is a worldwide zoonosis caused by spirochetes from the genus Leptospira. In the present study, a visual and rapid method for detecting Leptospira interrogans was developed based on multiple cross-displacement amplification (MCDA) and nanoparticle-based lateral flow biosensor (LFB). A set of 10 primers was specifically designed to recognize 10 regions of the lipL 41 gene of L. interrogans. The MCDA reaction was optimized at 64°C for only 40 min, and the amplification products were directly applied to the biosensor. The entire process, including DNA extraction (25 min), MCDA reaction (40 min), and result interpretation (∼2 min), could be completed within 70 min. Amplification products were detectable from as few as 10 genomic equivalents per reaction of pure L. interrogans DNA. No cross-reaction with nonpathogenic Leptospira or other bacteria was observed. The MCDA-LFB method established in the current report is suitable for the rapid screening of L. interrogans in clinical, animal, and environmental samples.