Isolation and characterization of two European strains of Ehrlichia phagocytophila of equine origin.
Abstract: We report the isolation and partial genetic characterization of two equine strains of granulocytic Ehrlichia of the genogroup Ehrlichia phagocytophila. Frozen whole-blood samples from two Swedish horses with laboratory-verified granulocytic ehrlichiosis were inoculated into HL-60 cell cultures. Granulocytic Ehrlichia was isolated and propagated from both horses. DNA extracts from the respective strains were amplified by PCR using primers directed towards the 16S rRNA gene, the groESL heat shock operon gene, and the ank gene. The amplified gene fragments were sequenced and compared to known sequences in the GenBank database. With respect to the 16S rRNA gene, the groESL gene, and the ank gene, the DNA sequences of the two equine Ehrlichia isolates were identical to sequences found in isolates from clinical cases of granulocytic ehrlichiosis in humans and domestic animals in Sweden. However, compared to amplified DNA from an American Ehrlichia strain of the E. phagocytophila genogroup, differences were found in the groESL gene and ank gene sequences.
Publication Date: 2002-03-05 PubMed ID: 11874874PubMed Central: PMC119922DOI: 10.1128/cdli.9.2.341-343.2002Google Scholar: Lookup
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Summary
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The research focuses on isolating and genetically defining two strains of granulocytic Ehrlichia, cells causing horse diseases, from the genogroup Ehrlichia phagocytophila in Sweden. It was found that their DNA makeup matches those of the strains on humans and other domestic animals from the same region.
Isolation and Propagation
- The study was performed using frozen blood samples from two horses diagnosed with granulocytic ehrlichiosis, a disease caused by the granulocytic Ehrlichia.
- These samples were inoculated into HL-60 cell cultures, a type of human promyelocytic leukemia cell line, to isolate and propagate the Ehrlichia.
Genetic Characterization
- The researchers used a technique known as PCR (polymerase chain reaction) to amplify the DNA extracts from the strains.
- They used specific primers to amplify the 16S rRNA gene, the groESL heat shock operon gene, and the ank gene, which are key to the Ehrlichia’s makeup and functionality.
- They then compared these amplified gene fragments to known sequences listed in the GenBank database, which is a repository of all publicly available genetic sequences.
Comparisons and Discoveries
- The study found that the DNA sequences of the isolated equine Ehrlichia strains were identical to the sequences in Ehrlichia strains from humans and other domestic animals in Sweden who had been diagnosed with granulocytic ehrlichiosis.
- This discovery suggested that the strains in horses and those in humans and other domestic animals in Sweden were shared or common.
- On the other hand, when the researchers compared these strains to an American Ehrlichia strain from the E. phagocytophila genogroup, they found differences in the groESL gene and ank gene sequences. This points to geographical or evolutionary differences in the strains of Ehrlichia in different regions.
Cite This Article
APA
Bjöersdorff A, Bagert B, Massung RF, Gusa A, Eliasson I.
(2002).
Isolation and characterization of two European strains of Ehrlichia phagocytophila of equine origin.
Clin Diagn Lab Immunol, 9(2), 341-343.
https://doi.org/10.1128/cdli.9.2.341-343.2002 Publication
Researcher Affiliations
- Department of Clinical Microbiology, Kalmar County Hospital, SE-391 85 Kalmar, Sweden. anneli.bjoersdorff@ltkalmar.se
MeSH Terms
- Animals
- Antigens, Bacterial / genetics
- Bacterial Proteins / genetics
- Chaperonins / genetics
- Ehrlichia / classification
- Ehrlichia / genetics
- Ehrlichia / isolation & purification
- Ehrlichiosis / diagnosis
- Ehrlichiosis / veterinary
- HL-60 Cells
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horses
- Humans
- RNA, Bacterial / analysis
- RNA, Ribosomal, 16S / analysis
- Sweden
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Citations
This article has been cited 7 times.- Rule EK, Boyle AG, Stefanovski D, Anis E, Linton J, Lorello O. Transfer of naturally acquired specific passive immunity against Anaplasma phagocytophilum in foals in Southeastern Pennsylvania and Northern Maryland. J Vet Intern Med 2023 Sep-Oct;37(5):1889-1892.
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