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Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay.

Publication ,  Journal Article
Gusa, AA; Buller, RS; Storch, GA; Huycke, MM; Machado, LJ; Slater, LN; Stockham, SL; Massung, RF
Published in: J Clin Microbiol
November 2001

PCR was used to amplify a 537-bp region of an Ehrlichia ewingii gene encoding a homologue of the 28-kDa major antigenic protein (P28) of Ehrlichia chaffeensis. The E. ewingii p28 gene homologue was amplified from DNA extracted from whole blood obtained from four humans and one canine with confirmed cases of infection. Sequencing of the PCR products (505 bp) revealed a partial gene with homology to outer membrane protein genes from Ehrlichia and Cowdria spp.: p30 of Ehrlichia canis (< or =71.3%), p28 of E. chaffeensis (< or =68.3%), and map1 of Cowdria ruminantium (67.3%). The peptide sequence of the E. ewingii partial gene product was deduced (168 amino acids) and the antigenicity profile was analyzed, revealing a hydrophilic protein with < or =69.1% identity to P28 of E. chaffeensis, < or =67.3% identity to P30 of E. canis, and < or =63.1% identity to MAP1 of C. ruminantium. Primers were selected from the E. ewingii p28 sequence and used to develop a species-specific PCR diagnostic assay. The p28 PCR assay amplified the expected 215-bp product from DNA that was extracted from EDTA-treated blood from each of the confirmed E. ewingii infections that were available. The assay did not produce PCR products with DNA extracted from E. chaffeensis-, E. canis-, or E. phagocytophila-infected samples, confirming the specificity of the p28 assay for E. ewingii. The sensitivity of the E. ewingii-specific PCR assay was evaluated and determined to detect as few as 38 copies of the p28 gene.

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Published In

J Clin Microbiol

DOI

ISSN

0095-1137

Publication Date

November 2001

Volume

39

Issue

11

Start / End Page

3871 / 3876

Location

United States

Related Subject Headings

  • Species Specificity
  • Sequence Analysis, DNA
  • Sensitivity and Specificity
  • Polymerase Chain Reaction
  • Molecular Sequence Data
  • Microbiology
  • Humans
  • Ehrlichiosis
  • Ehrlichia
  • Dogs
 

Citation

APA
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ICMJE
MLA
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Gusa, A. A., Buller, R. S., Storch, G. A., Huycke, M. M., Machado, L. J., Slater, L. N., … Massung, R. F. (2001). Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay. J Clin Microbiol, 39(11), 3871–3876. https://doi.org/10.1128/JCM.39.11.3871-3876.2001
Gusa, A. A., R. S. Buller, G. A. Storch, M. M. Huycke, L. J. Machado, L. N. Slater, S. L. Stockham, and R. F. Massung. “Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay.J Clin Microbiol 39, no. 11 (November 2001): 3871–76. https://doi.org/10.1128/JCM.39.11.3871-3876.2001.
Gusa AA, Buller RS, Storch GA, Huycke MM, Machado LJ, Slater LN, et al. Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay. J Clin Microbiol. 2001 Nov;39(11):3871–6.
Gusa, A. A., et al. “Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay.J Clin Microbiol, vol. 39, no. 11, Nov. 2001, pp. 3871–76. Pubmed, doi:10.1128/JCM.39.11.3871-3876.2001.
Gusa AA, Buller RS, Storch GA, Huycke MM, Machado LJ, Slater LN, Stockham SL, Massung RF. Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay. J Clin Microbiol. 2001 Nov;39(11):3871–3876.

Published In

J Clin Microbiol

DOI

ISSN

0095-1137

Publication Date

November 2001

Volume

39

Issue

11

Start / End Page

3871 / 3876

Location

United States

Related Subject Headings

  • Species Specificity
  • Sequence Analysis, DNA
  • Sensitivity and Specificity
  • Polymerase Chain Reaction
  • Molecular Sequence Data
  • Microbiology
  • Humans
  • Ehrlichiosis
  • Ehrlichia
  • Dogs