PCR detection of acute Ehrlichia canis infection in dogs

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

Research output: Contribution to journalArticle

60 Scopus citations

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
DOIs
StatePublished - Jan 1 1996

ASJC Scopus subject areas

  • veterinary(all)

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    McBride, J. W., Corstvet, R. E., Gaunt, S. D., Chinsangaram, J., Akita, G. Y., & Osburn, B. (1996). PCR detection of acute Ehrlichia canis infection in dogs. Journal of Veterinary Diagnostic Investigation, 8(4), 441-447. https://doi.org/10.1177/104063879600800406