J Bacteriol Virol.  2014 Mar;44(1):59-66. 10.4167/jbv.2014.44.1.59.

Identification of Leptospira species of Korean Isolates using Phylogenetic Analysis of Polymerase Chain Reaction-amplified 16S rDNA and LipL32 Genes

Affiliations
  • 1Department of Microbiology, College of Medicine, Konkuk University, Seoul, Korea. wjjang@kku.ac.kr
  • 2Institute of Glocal Disease Control, College of Medicine, Konkuk University, Seoul, Korea.
  • 3Department of Microbiology, College of Medicine, Hallym University, Chunchon, Kangwon-do, Korea.

Abstract

In this study, we selected only serologically identified 15 Leptosira interrogans isolates in the past and analyzed and identified them by using molecular method. The partial 16S rDNA and LipL32 genes were amplified from the bacteria by polymerase chain reaction (PCR) and sequenced. Sizes of the PCR products were 529 bp and 819 bp respectively and analysis of the nucleotide sequence of 16S rDNA and LipL32 genes showed that 14 out the 15 Leptospira showed 99.4~100% and 99.2~99.9% similarity respectively to those of L. interrogans lai and one isolate named HS-7 showed 100% and 100% similarity to L. interrogans canicola. The phylogenetic tree based on the 16S rDNA and LipL32 genes obtained the study revealed that 14 of the Leptospira composed a cluster distinct to that of L. interrogans lai and HS-7 composed to L. interrogans canicola.

Keyword

Leptospira interrogans; Serovar; Genospecies

MeSH Terms

Bacteria
Base Sequence
DNA, Ribosomal*
Leptospira interrogans
Leptospira*
Polymerase Chain Reaction
DNA, Ribosomal

Figure

  • Figure 1. Dendrogram representing phylogenetic relationships between partial 16S rDNA sequences (the size of about 323 bp) of reference Leptospira strains and 16S rDNA PCR products of isolates. The phylogram was generated by neighbor-joining analysis with 1,000 bootstrapped replicates.

  • Figure 2. Dendrogram representing phylogenetic relationships between partial LipL 32 sequences (the size of about 727 bp) of reference Leptospira strains and LipL 32 PCR products of isolates. The phylogram was generated by neighbor-joining analysis with 1,000 bootstrapped replicates.


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