J Bacteriol Virol.  2009 Dec;39(4):317-327. 10.4167/jbv.2009.39.4.317.

Detection and Identification of the Spotted Fever Group Rickettsial Agents from Haemaphysalis Ticks in Jeju Island, Korea

• Moon BC
• Jeong JH
• Choi YJ
• Kim JE
• Seo HJ
• Shin EH
• Song BG
• Lee HI
• Lee SH
• Park KH
• Jang WJ

• KMID: 2168543
• DOI: http://doi.org/10.4167/jbv.2009.39.4.317

Abstract

This study investigated the presence of nucleic acids of various Rickettsial agents in ticks collected in Jeju Island, Korea from June 2007 to August 2008, through the nested polymerase chain reaction (PCR) and sequencing analysis of partial citrate synthase (gltA), Rickettsial outer membrane protein B (ompB), and 17-kDa genes. Examination of the 1,584 ticks showed that the subspecies distribution of Haemaphysalis longicornis was 99.81% (n=1,581) and H. flava was 0.19% (n=3). A total 224 out of 250 pools from one to 15 ticks were found to be positive in ompB-PCR assay (minimal infection rate 141 ticks/1,000 tested). From the positive samples, 26 were analyzed by gltA- and 17-kDa-PCR assays. The nucleotide sequences of the ompB- and gltA-PCR products showed a high degree of similarity with those of the Rickettsia japonica (98.7~99.2% and 98.7~99.3%, n=25) and R. monacensis (99% and 99.7%, n=1). However, analysis of the nucleotide sequences of the 17-kDa-PCR amplicons showed that the sequences of the 25 PCR amplicons were more close to R. marmionii (99.4~100%) than R. japonica (98.6~99.1%). These findings suggest that various rickettsial diseases could be transmitted via the bite of tick vectors in Jeju Island, Korea.

Keyword

Rickettsial agents; Tick; Polymerase chain reaction; gltA; ompB; 17-kDa gene

MeSH Terms

Base Sequence
Bites and Stings
Citrate (si)-Synthase
Fever
Korea
Membrane Proteins
Nucleic Acids
Polymerase Chain Reaction
Receptors, Fc
Rickettsia
Ticks
Citrate (si)-Synthase
Membrane Proteins
Nucleic Acids
Receptors, Fc

Figure

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  Figure 1. Map of Jeju Island, Korea showing the geographic distribution of ticks collected from 2007 to 2008. Dots indicate the collection area of the H. longicornis and squares indicate the collection area of H. longicornis and H. flava.

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  Figure 2. Electrophoresis analysis on 1.2% agarose gel of DNAs amplified by PCR targeted ompB (panel A), gltA (panel B), and the 17-kDa (panel C) antigen gene. (A) The size of amplified ompB product was about 407 bp. Lane 1, positive control (R. japonica); lane 2~4, each number of the amplified ompB products. (B) The size of amplified gltA product was about 330 bp. Lane 1, positive control (R. japonica); lane 2~3, each number of the amplified gltA products. (C) The size of amplified 17-kDa product was about 360 bp. Lane 1, positive control (R. japonica); lane 2~3, each number of the amplified 17-kDa products. Lane M, 100 bp DNA ladder; lane N, negative control. The number on the left indicates the molecular size (in base pairs) of the amplified PCR products.

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  Figure 3. Dendrogram representing phylogenetic relationships between partial ompB gene sequences (the size of about 407 bp) of various Rickettsial strain and PCR-amplified ompB products from tick. Phylograms were generated by neighbor-joining analysis with 1,000 bootstrapped replicates.

• 

  Figure 4. Dendrogram representing phylogenetic relationships between partial gltA gene sequences (the size of about 330 bp) of various Rickettsial strains and PCR-amplified gltA products from tick. Phylograms were generated by neighbor-joining analysis with 1,000 bootstrapped replicates.

• 

  Figure 5. Dendrogram representing phylogenetic relationships between partial 17-kDa gene sequences (the size of about 360 bp) of various Rickettsial strain and PCR-amplified 17-kDa products from tick. Phylograms were generated by neighbor-joining analysis with 1,000 bootstrapped replicates.

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