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Leptospirosis is an emerging zoonotic disease endemic in Kerala and close monitoringof emerging serovars is essential to adopt appropriate control strategies. Multi-Locus SequenceTyping (MLST) was reported to be less expensive compared to other cumbersome methods likewhole genome sequencing. The present study was conducted to obtain isolates of Leptospira frominfected animals and rats and for the identification of serovars using MLST. A total of 205 bloodsamples (dog, cat, cattle, goat), 43 urine samples (dog, cattle) and post-mortem kidney samplesfrom various animals such as dog (n=12), cattle (n=2) and rat (n=25) were collected and subjectedto polymerase chain reaction (PCR) using G1/G2 primers to identify the pathogenic Leptospira.Fifteen samples were found to be positive, these samples when inoculated in the Ellinghausen-McCullough-Johnson-Harris (EMJH) semi-solid medium to obtain ten isolates. These ten isolateswere further subjected to secY, icdA and GyraseB PCR and sequenced. The obtained sequenceswere analysed using BLAST and were fed into specified MLST database of Leptospira scheme-3,the allelic profile and sequence type were generated. The MLST results obtained in the studyindicated that the isolates S24 and S33 belonged to serovar Canicola, S40 and 47 were Sejroeand S19, S27, S55, S69 and S71 were Bataviae, Autumnalis, Pomona, Icterohaemorraghiae andAustralis, respectively. It was concluded that MLST is a convenient method for identifying leptospiralserovars.

Multi-locus sequence typing for species/serovar identification of clinical isolates of Leptospira spp