Characterization of Borrelia Burgdorferi Isolated from Ixodes Persulacatus and Ixodes Ovatus Ticks in Japan

Nakao, Minoru; Miyamoto, Kenji; Uchikawa, Kimito; Fujita, Hiromi

doi:10.4269/ajtmh.1992.47.505

Cited by 65 publications

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“…Interestingly, all the isolates from I. ovatus were homogeneous with respect to the molecular size of Osp A (25). On the other hand, that of the isolates from I. persulcatus was heterogeneous (14,25). Recently, we proposed B. japonica isolated from I. ovatus in Japan on the basis of genomic DNA hybridization analysis and RFLP analyses of the fla and 16S rRNA genes (19).…”

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“…Japanese isolates were characterized bacteriologically based on protein profiles (25), plasmid profiles (32), restriction fragment length polymorphism (RFLP) analysis in 23S rRNA gene, 16S rRNA gene (12,13,15,19) and fla (19) and reactivities with MAbs against Osp A and flagellin (25). Interestingly, all the isolates from I. ovatus were homogeneous with respect to the molecular size of Osp A (25). On the other hand, that of the isolates from I. persulcatus was heterogeneous (14,25).…”

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Polymerase Chain Reaction Analysis of Borrelia Species Isolated in Japan

Kawabata

Tashibu

Yamada

et al. 1994

Microbiology and Immunology

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Primer reactivities of 25 Borrelia burgdorferi sensu lato isolates from the ticks, Ixodes persulcatus and I. ovatus, in Japan and 10 isolates in Europe and North America were investigated. The methods used in this study were the polymerase chain reaction (PCR) on the flagellin structural gene (fla), the outer surface protein A gene (osp A) and the outer surface protein B gene (osp B), and the restriction fragment length polymorphism (RFLP) analysis of PCR products from osp A and osp B. The flagellin PCR primer set reacted with all the Borrelia strains tested. Four genospecies, B. burgdorferi sensu stricto, B. garinii, B. afzelii and B. japonica, were differentiated by PCR using osp A and osp B primers combined with RFLP analysis. Some Japanese isolates from I. persulcatus were identified as B. garinii or B. afzelii. The other isolates from I. persulcatus did not fit in any of the 4 genospecies.These results suggested that Japanese isolates from I. persulcatus are highly heterogeneous in their osp A and osp B structures. Furthermore, PCR primers targetingfla are applicable to the gene diagnosis for Lyme disease in Japan, and osp A and osp B primers can be used to classify B. burgdorferi sensu lato isolates into genospecies by PCR and RFLP analyses.

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Polymerase Chain Reaction Analysis of Borrelia Species Isolated in Japan

Kawabata

Tashibu

Yamada

et al. 1994

Microbiology and Immunology

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“…The aetiological agent of Lyme disease, Borrelia burgdorferi sensu lato, is transmitted mainly by ticks of the Ixodes ricinus complex in North America and Europe [4,5] and by I. persulcatus and I. ovatus ticks in the countries of Far East Asia [6][7][8]. Although a human case of Lyme disease had been reported in Taiwan [9] and Borrelia spirochaetes were also isolated from rodents in the Taiwan area [10], the genetic diversity of spirochaetes as well as the tick vectors responsible for transmission in Taiwan need to be established.…”

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Genetic analysis of the outer surface protein C gene of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) isolated from rodents in Taiwan

Shih

Chao

2002

Journal of Medical Microbiology

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The outer surface protein C gene (ospC) of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) was analysed for the first time in Taiwan. The genetic identities of these Taiwan isolates (TWKM1-7) were determined by restriction fragment length polymorphism (RFLP) analysis and sequence similarities of the PCR-amplified ospC gene amplicons. After cleavage by nuclease Dra I, differential fragment patterns of PCRamplified ospC DNA in relation to different genospecies of Lyme disease spirochaetes were observed and all of these Taiwan isolates were genetically aff iliated to the genospecies of B. burgdorferi sensu stricto. The phylogenetic analysis on the sequence similarity of these Taiwan isolates revealed a highly homogeneous genotype, ranging from 99.3% to 100%, within the genospecies of B. burgdorferi sensu stricto and was distinguished from other genospecies of Borrelia isolates. The sequence similarity analysis also revealed the high sequence variability of the ospC gene among Borrelia strains that belong to the same genospecies but were isolated from different biological and geographical sources. Thus, these results provide the first investigation on the genetic identity of the ospC gene of these Taiwan isolates and show that these Taiwan isolates were closely related genetically to the genospecies of B. burgdorferi sensu stricto.

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“…The major species distributed in Japan is Borrelia garinii, and the most prevalent areas of distribution are Nagano and Hokkaido (18,20). Borrelia burgdorferi sensu lato have several major outer surface proteins (Osps), and the genes for OspA and OspB are located in tandem on a large linear extra-chromosomal DNA molecule that is 49-kb in size (3,15).…”

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Genetic Diversity and the Absence of Regional Differences of Borrelia garinii as Demonstrated by ospA and ospB Gene Sequence Analysis

Yabuki

Nakao

Fukunaga

1999

Microbiology and Immunology

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Unfed adult Ixodes persulcatus ticks were collected from four locations of Nagano and Hokkaido in Japan. Infected Borrelia garinii were investigated by PCR-RFLP of the ospA and ospB gene sequences. The primer set amplified an approximately 1.6-kb DNA fragment (0.7-kb in some strains), and Bsrl, BstYI, or N/aIII digestion of the product resulted in six distinctively different PCR-RFLP groups and two independent borrelial strains. The representatives in each PCR-RFLP group and individuals from the borrelial strains were sequenced, and their deduced amino acid sequences were aligned. A neighbor-joining phylogenetic analysis showed that the B. garinii OspA or OspB sequences were each divided into three major clusters including isolates from both the Nagano and Hokkaido locations. There was no local difference in OspA/B sequences between Nagano and Hokkaido. The osp gene of Borrelia burgdorferi sensu lato is highly heterogeneous, and this was also confirmed by our sequence analysis. Some strains of the different PCR-RFLP groups had closely related OspA sequences, while the OspB sequences of these strains were quite different. These findings suggested intraspecies gene exchange and recombination events between the two genes in B. garinii.

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Characterization of Borrelia Burgdorferi Isolated from Ixodes Persulacatus and Ixodes Ovatus Ticks in Japan

Cited by 65 publications

References 0 publications

Polymerase Chain Reaction Analysis of Borrelia Species Isolated in Japan

Polymerase Chain Reaction Analysis of Borrelia Species Isolated in Japan

Genetic analysis of the outer surface protein C gene of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) isolated from rodents in Taiwan

Genetic Diversity and the Absence of Regional Differences of Borrelia garinii as Demonstrated by ospA and ospB Gene Sequence Analysis

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