Application of Multilocus Sequence Typing for the Characterization of Leptospira Strains in Malaysia.

Leptospirosis is a common zoonotic disease in tropical and subtropical countries. It is considered an emerging disease in Malaysia and is a notifiable disease. This study was conducted to characterize Malaysian isolates from human, animal and environmental samples via MLST and rrs2 sequencing in an attempt to develop a Malaysian genotypic database. An existing polymerase chain reaction (PCR)-based MLST scheme was performed to facilitate subsequent sequencing. Out of 46 extracted DNA, 36 had complete MLST profiles whereby all six genes were amplified and sequenced. Most of the pathogenic Leptospira genotypes with full MLST profiles were L. interrogans serogroup Bataviae (n = 17), followed by L. borgpetersenii serogroup Javanica (n = 9), L. interrogans serogroup Sejroe (n = 2), L. interrogans serogroup Australis (n = 2), L. kirschneri (n = 2), L. interrogans serogroup Grippotyphosa (n = 1) and L. interrogans serogroup Pyrogenes (n = 3). Two samples (R3_SER/17 and R4_SER/17) were not closely related with any of the reference strains. For the samples with incomplete MLST profiles, leptospiral speciation was conducted through rrs2 analysis, in which four samples were identified as L. borgpetersenii, five samples were closely related to L. kmetyi and one sample was known as L. yasudae. This study shows that molecular approaches that combine both MLST and rrs2 sequencing have great potential in the comprehensive characterization of pathogenic Leptospira because they can be performed directly from cultured and clinical samples.