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Journal of clinical microbiology1996; 34(9); 2170-2174; doi: 10.1128/jcm.34.9.2170-2174.1996

A 16S rRNA-based PCR assay for detection and identification of granulocytic Ehrlichia species in dogs, horses, and cattle.

Abstract: A PCR-based assay was developed for detecting DNA of granulocytic ehrlichiae in blood samples from dogs, horses, and cattle, Primers were designed from 16S rRNA sequence information to specifically amplify DNA from a newly identified Swedish Ehrlichia species. The 16S rRNA nucleotide sequence of this Swedish species differs in only two and three positions from the sequences of Ehrlichia phagocytophila and Ehrlichia equi, respectively, which were also amplified by this PCR system. For evaluation, PCR results were compared with microscopic examination of stained blood smears for the detection of granulocytes containing ehrlichiae (morulae). Thirty-four of 36 microscopically positive samples were also positive by PCR, and 6 microscopically negative samples were negative by PCR as well. Six samples, in which morulae-like structures had been seen, were negative by PCR, also at a lower annealing temperature and when a reamplification of the first PCR products was performed. The identities of the PCR products from some canine and equine isolates were verified by direct DNA sequencing and were found to be identical with the Ehrlichia sequence found in these animal species that had been obtained earlier. The sequences of a segment of approximately 600 nucleotides from two bovine isolates were identical to that of E. phagocytophila, whereas the sequence of another bovine isolate differed in two positions from that of E. phagocytophila and in three positions from the sequences of the canine and equine isolates. Serum samples were analyzed by indirect fluorescent-antibody testing. Seventy-three percent of the animals which were positive by microscopy and PCR also had positive antibody titers. However, it was not possible to rely on a single serological result for diagnosis of present infection. It was, therefore, concluded that PCR was the most reliable method, useful in the clinical laboratory for specific and early diagnosis of granulocytic ehrlichiosis in animals.
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Publication Date: 1996-09-01 PubMed ID: 8862579PubMed Central: PMC229211DOI: 10.1128/jcm.34.9.2170-2174.1996Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper details the development of a PCR-based assay for the detection of a specific type of bacteria in animals such as dogs, horses, and cattle. The assay is reliable and provides a useful and specific way for diagnosis of granulocytic ehrlichiosis in these animals.

Methodology

In this study, the researchers:

  • Developed a PCR-based assay to detect DNA of granulocytic ehrlichiae in blood samples from dogs, horses, and cattle.
  • Designed primers from 16S rRNA sequence information to specifically amplify DNA from a newly identified Swedish Ehrlichia species.
  • Compared the results of the PCR test with microscopic examinations of stained blood smears to detect granulocytes containing ehrlichiae.
  • Used indirect fluorescent-antibody testing to analyze serum samples.

Findings

The findings of the research include:

  • 34 of 36 microscopically positive samples were also PCR positive, implying a high concordance between the two methods.
  • The sequences of a segment of approximately 600 nucleotides from two bovine isolates were identical to that of E. phagocytophila, while the sequence of another bovine isolate differed in two positions from that of E. phagocytophila and in three positions from the sequences of the canine and equine isolates.
  • 73% of the animals which were positive by both microscopy and PCR also had positive antibody titers.

Conclusion

The study concluded that:

  • Due to its efficacy, the PCR test is the most reliable method for diagnosing granulocytic ehrlichiosis in animals.
  • While serological results were useful, a single serological result was insufficient for a definitive diagnosis of the condition.

Cite This Article

APA
Engvall EO, Pettersson B, Persson M, Artursson K, Johansson KE. (1996). A 16S rRNA-based PCR assay for detection and identification of granulocytic Ehrlichia species in dogs, horses, and cattle. J Clin Microbiol, 34(9), 2170-2174. https://doi.org/10.1128/jcm.34.9.2170-2174.1996

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 34
Issue: 9
Pages: 2170-2174

Researcher Affiliations

Engvall, E O
  • National Veterinary Institute, Uppsala, Sweden. eva.olsson@sva.se
Pettersson, B
    Persson, M
      Artursson, K
        Johansson, K E

          MeSH Terms

          • Animals
          • Cattle
          • Dogs
          • Ehrlichia / genetics
          • Ehrlichia / isolation & purification
          • Horses / microbiology
          • Leukocytes / microbiology
          • Molecular Sequence Data
          • Polymerase Chain Reaction / methods
          • RNA, Bacterial / analysis
          • RNA, Ribosomal, 16S / analysis

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