Detection and molecular characterisation of equine infectious anaemia virus from field outbreaks in Slovenia.
Abstract: In 2009, a surprisingly high number of animals seropositive for equine infectious anaemia virus (EIAV; 26 horses from 13 farms) were detected in Slovenia. Objective: To develop a polymerase chain reaction (PCR) assay for the detection of the proviral nucleic acid, to phylogenetically characterise the Slovenian EIAV strains and to investigate whether transmission in utero occurred. Methods: Cross-sectional clinical study. Methods: In total, 26 horses (including 2 foals and 4 pregnant mares) and 4 fetuses were examined in this study. A PCR assay using the EIAV F1 and EIAV R1 primers was designed and tested using genomic DNA extracted from 28 spleen samples, 18 whole blood samples and 17 peripheral blood leucocyte samples. Amplicons of 22 PCRs obtained from the spleen samples were subjected to direct DNA sequencing and phylogenetic analysis. Results: All spleen samples from 22 adult animals were positive for EIAV by PCR, whereas whole blood and the peripheral blood leucocyte samples were positive from only 4 animals. Spleen samples from foals and fetuses were negative by PCR. The Slovenian EIAV sequences could be mapped to 9 different branches of the phylogenetic tree. Conclusions: The PCR was able to detect different EIAV strains from spleen samples of seropositive animals detected in Slovenia. Phylogenetic analysis revealed high genetic diversity of the EIAV strains detected in Slovenia, with their closest relatives being European strains. In utero transmission in pregnant mares did not occur.
© 2013 EVJ Ltd.
Publication Date: 2013-09-09 PubMed ID: 23834226DOI: 10.1111/evj.12138Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research is about the detection and profiling of equine infectious anaemia virus (EIAV) that was surprisingly high among seropositive horses in Slovenia in 2009. A PCR assay was developed for detection and phylogenetic characterization of these strains, concluding a high genetic diversity among them and no evidence of in utero transmission in the affected mares.
Objective
The primary aim of the research was:
- Development of a polymerase chain reaction (PCR) assay for the detection of EIAV
- Characterize EIAV in Slovenia phylogenetically
- Investigate the possibility of in utero transmission
Methods
The methods involved the following process:
- A total of 26 horses were examined, which included 2 foals and 4 pregnant mares, along with 4 fetuses.
- The researchers designed a PCR assay using erythrocyte EIAV F1 and R1 primers. The genomic DNA was extracted from 28 spleen samples, 18 whole blood samples, and 17 peripheral blood leucocyte samples and tested.
- The amplicons derived from 22 PCR results from the spleen samples were put through direct DNA sequencing and a phylogenetic analysis.
Results
The results were as follows:
- All spleen samples from the 22 adult animals showed positive results for EIAV when tested using the PCR assay.
- The whole blood and peripheral blood leucocyte samples showed positive results for EIAV for only 4 animals.
- The spleen samples from foals and fetuses were negative for EIAV.
- The analysed Slovenian EIAV sequences were seen to branch into 9 different sections in the phylogenetic tree.
Conclusions
The conclusion was drawn based on the methods and results:
- PCR proves to be useful in detecting different strains of EIAV in spleen samples from seropositive animals found in Slovenia.
- The EIAV strains detected in Slovenia presented a high level of genetic diversity and were closely related to strains found in European horses.
- The study found no occurrence of in utero transmission in the pregnant mares that were tested.
Cite This Article
APA
Kuhar U, Završnik J, Toplak I, Malovrh T.
(2013).
Detection and molecular characterisation of equine infectious anaemia virus from field outbreaks in Slovenia.
Equine Vet J, 46(3), 386-391.
https://doi.org/10.1111/evj.12138 Publication
Researcher Affiliations
- Veterinary Faculty, Institute for Microbiology and Parasitology, Virology Unit, University of Ljubljana, Slovenia.
MeSH Terms
- Animals
- Disease Outbreaks / veterinary
- Equine Infectious Anemia / epidemiology
- Equine Infectious Anemia / virology
- Female
- Genetic Variation
- Horses
- Infectious Anemia Virus, Equine / genetics
- Infectious Anemia Virus, Equine / isolation & purification
- Male
- Phylogeny
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Slovenia / epidemiology
Citations
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