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Immunohistochemical diagnosis of eastern equine encephalomyelitis.
Abstract: An immunohistochemical (IHC) assay was developed for the detection of eastern equine encephalomyelitis (EEE) virus antigen in formalin-fixed, paraffin-embedded tissues. All cases of EEE diagnosed at the Michigan State University Animal Health Diagnostic Laboratory from 1991 through 1994 were evaluated. The diagnosis was based on histopathologic examination of the brain and confirmatory virus, isolation. Sections of cerebrum from 26 equids and 5 birds were assessed by IHC. Histologically normal brain tissues from 2 horses and 1 pheasant and brain tissues from 2 cases of equine neurologic disease with diagnoses other than EEE served as negative controls. The IHC assay was based on standard streptavidin-biotin technology, using a commercially available kit and a monospecific polyclonal primary antibody preparation derived from murine ascitic fluid. Nineteen of 20 equids and all 5 birds positive by histopathology and virus isolation were positive for EEE virus antigen by IHC. Three equids with histologic lesions compatible with a diagnosis of EEE but negative by virus isolation also were negative for virus antigen by IHC. In 3 other equids, histopathology and IHC were positive for EEE, but virus isolation was not attempted because of contamination of the brain specimen. The IHC assay of formalin-fixed, paraffin-embedded brain tissues for EEE virus antigen is a rapid, effective test for confirming a histopathologic diagnosis of EEE, and assay results correlate well with virus isolation results.
Publication Date: 1996-04-01 PubMed ID: 8744735DOI: 10.1177/104063879600800203Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
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.
This study details the development and application of an immunohistochemical (IHC) assay that detects the eastern equine encephalomyelitis (EEE) virus antigen in fixed, processed tissues. The assay was tested on brain tissue samples from horses and birds and was found to be an effective method of confirming a diagnosis of EEE.
Methodology
- The researchers developed an IHC assay for detecting the EEE virus antigen. This process uses formalin-fixed, paraffin-embedded tissues, thus enabling in-depth study of disease processes at the cellular level.
- Brain tissue sections from 26 horses and 5 birds, which all had previously been diagnosed with EEE at the Michigan State University Animal Health Diagnostic Laboratory between 1991-1994, were evaluated using the IHC assay.
- In order to verify the reliability of the test, the researchers also used it on control tissues. These included histologically normal brain tissues from 2 horses and 1 bird, as well as from 2 cases of equine neurological disease with diagnoses other than EEE.
Process of IHC Assay
- The IHC assay was conducted using a commercially available kit and a monospecific primary antibody preparation produced from murine ascitic fluid.
- This method relies on standard streptavidin-biotin technology, which enables specific and robust detection of target antigens in tissue sections.
Results
- Of the cases confirmed positive for EEE through histopathology and virus isolation, 95% (19 out of 20) of the horse cases and 100% of the bird cases were found to be positive for EEE virus antigen through the IHC assay.
- Some equine cases that had histologic lesions compatible with EEE but were negative by virus isolation were also found to be negative by the IHC assay. This finding indicates that the IHC assay accurately correlates with virus isolation results.
- For three horses, despite contamination of the brain specimen preventing virus isolation, both histopathology and IHC were positive for EEE. This suggests the IHC may be valuable in cases where other testing methods are not feasible.
Conclusion
- The results of the research show that the IHC assay provides a quick and effective method of corroborating a histopathological diagnosis of EEE.
- The results obtained from the IHC assay align well with the outcomes of virus isolation, demonstrating the assay’s reliability and accuracy.
Cite This Article
APA
Patterson JS, Maes RK, Mullaney TP, Benson CL.
(1996).
Immunohistochemical diagnosis of eastern equine encephalomyelitis.
J Vet Diagn Invest, 8(2), 156-160.
https://doi.org/10.1177/104063879600800203 Publication
Researcher Affiliations
- Animal Health Diagnostic Laboratory, College of Veterinary Medicine, Michigan State University, East Lansing 48824, USA.
MeSH Terms
- Animals
- Bird Diseases
- Birds
- Brain / pathology
- Brain / virology
- Encephalitis Virus, Eastern Equine / isolation & purification
- Encephalomyelitis, Equine / pathology
- Encephalomyelitis, Equine / veterinary
- Equidae
- Horse Diseases
- Horses
- Immunohistochemistry / methods
Citations
This article has been cited 3 times.- Williams JA, Long SY, Zeng X, Kuehl K, Babka AM, Davis NM, Liu J, Trefry JC, Daye S, Facemire PR, Iversen PL, Bavari S, Pitt ML, Nasar F. Eastern equine encephalitis virus rapidly infects and disseminates in the brain and spinal cord of cynomolgus macaques following aerosol challenge. PLoS Negl Trop Dis 2022 May;16(5):e0010081.
- Montalvo M, Ayoub D, McGary M, Byrd K, Mahmoud L, Mermel L, Thompson B, Wendell L. Eastern Equine Encephalitis: Case Series in Southern New England and Review of the Literature. Neurol Clin Pract 2021 Oct;11(5):e714-e721.
- Vogel P, Kell WM, Fritz DL, Parker MD, Schoepp RJ. Early events in the pathogenesis of eastern equine encephalitis virus in mice. Am J Pathol 2005 Jan;166(1):159-71.
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