Isolation and identification of Leptospira species from bovines by rpoB and LipL41 genes based phylogenetic analysis

摘要

In this study, in the beginning 582 serum samples were subjected to Microscopic Agglutination Test (MAT) with eight different serovars prevalent in the region to know the seroprevalence ofLeptospirain bovines in Karnataka, India. Based on the findings of the MAT, different samples like blood, urine, aborted materials and uterine discharge collected from the MAT positive animals were used for isolation and genomic detection by conventional PCR targeting twolipL32 and seqYgenes using specific primers. Out of the 163 MAT positive samples screened 12 samples (including three isolates) were found positive in PCR. Subsequently, to identify the different species prevalent in the geographical region the PCR positive samples were subjected torpoBandLipL41 gene amplification. and nucleotide sequence analysis ofrpo B, it was found that all the samples were belonging toL. interrogansspecies with overlapping/superimposingL. interrogansandL.borgpeterseniispecies. Further, theLipL41gene sequence phylogenetic analysis differentiated these two species clearly. Therefore, it can be concluded thatLipL41gene based phylogenic analysis besidesrpoBgene can be effectively utilized to identify differentLeptospiraspecies in a geographical niche including the identification of intermediate species. This is first of its kind in India usingLipL41gene based phylogenetic analysis forLeptospiraspecies identification in limited number of samples from bovines, hence the same can be explored on a larger geographical area with more number of samples and even to identify the prevalence or presence of intermediate species in different geographical locations.