Identification of Borrelia burgdorferi Isolated in Korea Using Outer Surface Protein A (OspA) Serotyping System

Abstract

Two characteristic strains (935T, 934U) of B. burgdorferi isolated from Ixodes persulcatus and a wild rodent (Apodemus agrarius) in Korea were selected and analyzed by an immunoblot method using the monoclonal antibodies directed to different epitopes of outer surface protein A (OspA). The reactive pattern of strain 934U with these monoclonal antibodies was identical to that of strains belonging to B. afzelii and that of strain 935T was different from other isolates. Monoclonal antibody (5TEE3) which is specific to strain 935T did not react with any other Western and Japanese isolates. So, it was suggested that there exist at least two groups of B. burgdorferi in Korea. One could be classified as B. afzelii and the other is a divergent group from three known species of B. burgdorferi sensu stricto, B. garinii and B. afzelii.

Genetic Analysis of Borrelia burgdorferi Sensu Lato in Korea Using Genomic Hybridization and 16S rRNA Gene Sequence Determination

“…3). On the basis of this result and previous OspA antigenic analyses (14,15), we thought that these two strains were genetically and antigenically related to B. afzelii.…”

“…However, 935T strain is located at a distinct position in the phylogenetic tree with a closer relation to B. garinii than to other species. In the case of antigenic analysis of 935T using an outer surface protein A (OspA)-monoclonal antibodies panel, the reactivity of 935T was rather distinct from that of other members of the three species (15). However, considering the antigenic heterogeneity of B. garinii (33) and a long common branch with B. garinii in phylogenetic tree, 935T strain might be in Table 2.…”

“…In these results, 934U and 17Y strains were tightly clustered with R-IP3 strain. So, in addition to the results of previous antigenic analyses (14,15), these two strains can be classified into species B. afzelii. However, 935T strain is located at a distinct position in the phylogenetic tree with a closer relation to B. garinii than to other species.…”

“…of Shizuoka, Japan), and HP3 was provided by M. Mori (National Institute of Health in Japan). The morphological and antigenic features of Korean isolates were described previously (14,15 DNA was extracted from B. burgdorferi as described by Le Fabvre et al (18). For the cloning of 16S rDNA, 100 ng of genomic DNA was subjected to polymerase chain reaction (PCR) in a mixture (50 il) of 10 mM Tris-HC1 (pH 8.3), 50 mM KC1, 1.5 mM MgC12, 0.2 mM of dNTP mixture, 0.5 lim each forward and reverse primers and 2.5 U Taq polymerase (AmpliTaq, Perkin-Elmer Cetus, U.S.A.).…”

“…Previously, we reported two antigenically distinct groups of B. burgdorferi sensu lato isolated in Korea using outer surface protein A (OspA) antigenicity (14,15). In this report, we analyzed these Korean isolates using RFLP patterns of rRNA gene, and phylogenetic analysis through the sequence determination of 16S rRNA gene of each group.…”