Rapid MALDI-TOF mass spectrometry identification of Leptospira organisms

Leptospirosis is a worldwide deadly zoonotic disease. Accurate identification of the causative Leptospira spp. spirochetes ascertains the pathogenic status of the isolates, identifies potential source of infection and recognises outbreaks. Species identification is currently based on technically demanding, time and resources consuming serological and molecular methods. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) recently emerged as a first-line method for the accurate identification of bacteria, yet no data issued for Leptospira spp. We investigated the potential of MALDI-TOF-MS for the rapid identification of Leptospira isolates. Starting from a 10(5)organisms/mL suspension, MALDI-TOF-MS yielded an unique protein profile for each one of 19 Leptospira species reference isolates with a 100% reproducibility over 12 repeats, allowing to create a Leptopsira database. MALDI-TOF-MS further accurately identified 20/21 additional reference isolates representative of various serogroups at the species level as Leptospira interrogans (n=12), Leptospira kirschneri (n=5), Leptospira borgpetersenii (n=3), Leptospira noguchii (n=1) with identification score value of 2-2.5. Furthermore, six clinical isolates previously identified by rpoB sequencing, were correctly identified by MALDI-TOF-MS as L. interrogans (n=5) and L. borgpetersenii (n=1) with identification score value of 2-2.6. Identification was achieved in 40 min starting from the Leptospira suspension. MALDI-TOF-MS could complement serological and sequencing-based methods for the first line, rapid identification of Leptospira isolates in the clinical microbiology laboratory.