PCR detection of acute Ehrlichia canis infection in dogs

Jere McBride, Richard E. Corstvet, Stephen D. Gaunt, Jarasvech Chinsangaram, Geoffrey Y. Akita, Bennie I. Osburn

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

A polymerase chain reaction (PCR)-based detection assay that specifically detected Ehrlichia canis in dogs with acute infections was developed. A region of the 16S ribpsomal RNA gene of E. canis was targeted for PCR amplification and chemiluminescent hybridization (CH) with a complementary internal 287-base pair (bp) oligonucleotide probe. The CH improved the PCR assay sensitivity 1,000-fold as compared with visualization on ethidium bromide-stained agarose gels. The PCR assay with CH (PCR/CH) detected as little as 30 fg of E. canis genomic DNA, the equivalent of approximately 150 E. canis organisms. The 495-bp product defined by the specific primers was not detected when genomic DNA from E. platys, E. chaffeensis, E. risticii, and E. equi were used in the PCR/CH assay. The PCR/CH assay was tested with unfractionated blood samples collected from 9 dogs experimentally infected with E. canis. The PCR/CH assay had greater detection sensitivity than did cell culture isolation (CCI) from infected blood. PCR/CH detected E. canis 7 days prior to CCI in 4 of 6 experimentally infected dogs. The results obtained with the PCR/CH assay otherwise consistently matched the results obtained by CCI. This PCR/CH assay is a rapid, sensitive, and specific method for E. canis detection with sensitivity comparable to or exceeding that of CCI. A diagnosis of E. canis using this PCR/CH assay can be made in 2 days as compared with 1-4 weeks for CCI. The PCR/CH assay appears to be an acceptable alternative or complement to current diagnostic techniques.

Original languageEnglish (US)
Pages (from-to)441-447
Number of pages7
JournalJournal of Veterinary Diagnostic Investigation
Volume8
Issue number4
StatePublished - Oct 1996
Externally publishedYes

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Ehrlichia canis
Luminescent Measurements
polymerase chain reaction
hybridization
Dogs
Polymerase Chain Reaction
dogs
Infection
infection
Cell Separation
assays
cell culture
Cell Culture Techniques
Base Pairing
genomics
oligonucleotide probes
Equus
Ethidium
Oligonucleotide Probes
DNA

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

Cite this

McBride, J., Corstvet, R. E., Gaunt, S. D., Chinsangaram, J., Akita, G. Y., & Osburn, B. I. (1996). PCR detection of acute Ehrlichia canis infection in dogs. Journal of Veterinary Diagnostic Investigation, 8(4), 441-447.

PCR detection of acute Ehrlichia canis infection in dogs. / McBride, Jere; Corstvet, Richard E.; Gaunt, Stephen D.; Chinsangaram, Jarasvech; Akita, Geoffrey Y.; Osburn, Bennie I.

In: Journal of Veterinary Diagnostic Investigation, Vol. 8, No. 4, 10.1996, p. 441-447.

Research output: Contribution to journalArticle

McBride, J, Corstvet, RE, Gaunt, SD, Chinsangaram, J, Akita, GY & Osburn, BI 1996, 'PCR detection of acute Ehrlichia canis infection in dogs', Journal of Veterinary Diagnostic Investigation, vol. 8, no. 4, pp. 441-447.
McBride J, Corstvet RE, Gaunt SD, Chinsangaram J, Akita GY, Osburn BI. PCR detection of acute Ehrlichia canis infection in dogs. Journal of Veterinary Diagnostic Investigation. 1996 Oct;8(4):441-447.
McBride, Jere ; Corstvet, Richard E. ; Gaunt, Stephen D. ; Chinsangaram, Jarasvech ; Akita, Geoffrey Y. ; Osburn, Bennie I. / PCR detection of acute Ehrlichia canis infection in dogs. In: Journal of Veterinary Diagnostic Investigation. 1996 ; Vol. 8, No. 4. pp. 441-447.
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