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. 2002 Sep;9(5):1132-6.
doi: 10.1128/cdli.9.5.1132-1136.2002.

Determination of the nucleotide sequences of heat shock operon groESL and the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys for phylogenetic and diagnostic studies

Hisashi Inokuma  1 , Kaori Fujii, Masaru Okuda, Takafumi Onishi, Jean-Pierre Beaufils, Didier Raoult, Philippe Brouqui
Affiliations

Determination of the nucleotide sequences of heat shock operon groESL and the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys for phylogenetic and diagnostic studies

Hisashi Inokuma et al. Clin Diagn Lab Immunol. 2002 Sep.

Abstract

The 1,670-bp nucleotide sequence of the heat shock operon groESL and the 1,236-bp sequence of the citrate synthase gene (gltA) of Anaplasma (Ehrlichia) platys were determined. The topology of the groEL- and gltA-based phylogenetic tree was similar to that derived from 16S rRNA gene analyses with distances. Both groESL- and gltA-based PCRs specific to A. platys were also developed based upon the alignment data.

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Figures

FIG. 1.
FIG. 1.
Phylogenetic relationship of various Anaplasma, Ehrlichia, and Neorickettsia spp. based on the nucleotide sequences of gltA (a) and groESL (b) genes. The neighbor-joining method was used to construct the phylogenetic tree with the ClustalW program. The scale bar represents 10% divergence. The numbers at nodes are the proportions of 100 bootstrap resamplings that support the topology shown. The GenBank accession numbers of the groESL sequences used to construct the phylogenetic tree and aligned data are as follows: A. phagocytophila (strain HGE agent), AF165812; E. equi, AF172162; E. phagocytophila, U96729; A. marginale, AF165812; E. chaffeensis, L1 0917; E. canis, U96731; E. muris, AF210459; Ehrlichia sp. detected from I. ovatus, AB032711; Ehrlichia (Cowdria) ruminantium, U13638; Neorickettsia (Ehrlichia) risticii, U96732; Neorickettsia (Ehrlichia) sennetsu, U88092; Rickettsia prowazekii, Y15783; Bartonella henselae, U96734. The GenBank accession numbers of the gltA sequences used for comparative analysis are as follows: A. phagocytophila (strain HGE agent), AF304136; E. equi, AF304137; E. phagocytophila, AF304138; A. marginale, AF304139; A. centrale, AF304141; E. chaffeensis, AF304142; E. canis, AF304143; E. muris, AF304144; Ehrlichia sp. detected from I. ovatus, AF304145; E. (C.) ruminantium, AF304146; N. (E.) risticii, AF304147; N. (E.) sennetsu, AF304148: N. helminthoeca, AF304149; R. prowazekii, M17149; B. henselae, U78514.
FIG. 2.
FIG. 2.
The sensitivities of the A. platys-specific PCR based upon the groESL (lanes 1 to 6) and gltA (lanes 7 to 12) genes were evaluated. DNA equivalent to that from 375, 37.5, 3.75, 0.375, and 0.0375 infected platelets was used as a template for the amplicons demonstrated in lanes 1 and 7, 2 and 8, 3 and 9, 4 and 10, and 5 and 11, respectively. Distilled water was used in lanes 6 and 12 as a negative control. Lane M, 100-bp DNA ladder.
FIG. 3.
FIG. 3.
A. platys-specific PCRs based upon the groESL (A) and gltA (B) genes were evaluated for their specificity with DNA from A. platys strains from Somieres, France (lane 1), Okinawa, Japan (lane 2), and Venezuela (lane 3), and from A. marginale (lane 4), A. centrale (lane 5), A. phagocytophila strain HGE agent (lane 6), E. equi (lane 7), E. phagocytophila (lane 8), W. pipientis (lane 9), E. canis (lane 10), and N. helminthoeca (lane 11), with distilled water as a negative control (lane N). Positive bands (arrows) were observed only with DNA of the three strains of A. platys in both groESL- and gltA-based PCRs.
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References

    1. Allsopp, M. T. E. P., E. S. Visser, J. L. du Plessis, S. W. Vogel, and B. A. Allsopp. 1997. Different organisms associated with heartwater as shown by analysis of 16S ribosomal RNA gene sequences. Vet. Parasitol. 71:283-300. - PubMed
    1. Beaufils, J.-P., H. Inokuma, J. Martin-Granel, P. Jumelle, M. Barbault-Jumelle, and P. Brouqui. 2002. Anaplasma platys (Ehrlichia platys) infection in a dog in France: description of the case, and characterization of the agent. Rev. Med. Vet. 153:85-90.
    1. Chae, J.-S., J. E. Foley, J. S. Dumler, and J. E. Madigan. 2000. Comparison of the nucleotide sequence of 16S rRNA, 444 Ep-ank, and groESL heat shock operon genes in naturally occurring Ehrlichia equi and human granulocytic ehrlichiosis agent isolates from northern California. J. Clin. Microbiol. 38:1364-1369. - PMC - PubMed
    1. Chang, A. C. H., W. L. Chang, C. T. Lin, M. J. Pan, and S. C. Lee. 1996. Canine infectious cyclic thrombocytopenia found in Taiwan. J. Vet. Med. Sci. 58:473-476. - PubMed
    1. Chang, W. L., and M. J. Pan. 1996. Specific amplification of Ehrlichia platys DNA from blood specimen by two-step PCR. J. Clin. Microbiol. 34:3142-3146. - PMC - PubMed

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