[A western blot test for the serological diagnosis of equine infectious anemia].
Abstract: After electrophoretic separation in SDS-PAGE structural proteins of the virus of Equine Infectious Anemia (EIA) were easily blotted by the semi-dry-blotting method onto nitrocellulose filters. Strips of these filters were used for antibody demonstration, and positive reactions thereof were intensified by a biotin-avidin-peroxidase system. Sensitivity of this system was so high as to allow readable interpretation of bands up to the dilution of 1:6,400 of a strongly positive serum. Frequently this procedure allowed to make a firm diagnostic Western-Blot diagnosis on far weaker equine sera. Interpretation of results proved, however, difficult with sera, which formed one single band only. This observation, although of weak grade, had to be made in some 5% of sera stemming from horses with a certainly negative history of EIA. Consequently, we conclude, that a policy followed in the serodiagnostic Western-Blot of human AIDS should also be adopted for the interpretation of the EIA Western-Blot, namely to declare as positive merely horse sera which evidence more than one single band, whereof at least one band should represent a viral glycoprotein.
Publication Date: 1989-02-01 PubMed ID: 2538978
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- English Abstract
- Journal Article
Summary
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The research article discusses a sensitive Western Blot test method, which has been developed for diagnosing Equine Infectious Anemia (EIA) using the antibodies present in horse serum.
Methodology
- The researchers started by separating the structural proteins of the EIA virus using a method referred to as SDS-PAGE electrophoretic separation.
- These separated proteins were then transferred onto nitrocellulose filters using a semi-dry-blotted method.
- Strips of these filters were utilized for the detection of antibodies. Any positive reactions were intensified with a biotin-avidin-peroxidase system to increase the visibility of the results.
Sensitivity
- The sensitivity of this method allowed for the interpretation of bands up to a dilution of 1:6,400 in a strongly positive serum.
- This advanced sensitivity also enabled them to confirm an EIA diagnosis with weaker equine sera, showcasing the effectiveness of the testing procedure.
Results Interpretation
- Interpreting the results was challenging when the serum from the tested animal formed a single band only.
- This single-band scenario was noted in around 5% of the tested sera that came from horses with confirmed negative EIA history.
Recommendation
- Basing on the challenges related to the interpretation of single-band results, the researchers recommended an approach similar to the one used in the serodiagnostic Western-Blot of human AIDS.
- They suggested that horse sera should be declared positive only if they show more than a single band, where at least one band should represent a viral glycoprotein.
In conclusion, the researchers developed a sensitive method for diagnosing Equine Infectious Anemia using the Western Blot test. However, they noted challenges with interpreting single-band results and recommended an approach similar to the AIDS diagnostic procedure for more accurate results interpretation.
Cite This Article
APA
Rossmanith W, Horvath E.
(1989).
[A western blot test for the serological diagnosis of equine infectious anemia].
Zentralbl Veterinarmed B, 36(1), 49-56.
Publication
Researcher Affiliations
MeSH Terms
- Animals
- Blotting, Western
- Electrophoresis, Polyacrylamide Gel
- Equine Infectious Anemia / diagnosis
- Horses
- Infectious Anemia Virus, Equine / analysis
- Viral Proteins / analysis
Citations
This article has been cited 2 times.- Alvarez I, Gutierrez G, Ostlund E, Barrandeguy M, Trono K. Western blot assay using recombinant p26 antigen for detection of equine infectious anemia virus-specific antibodies.. Clin Vaccine Immunol 2007 Dec;14(12):1646-8.
- Langemeier JL, Cook SJ, Cook RF, Rushlow KE, Montelaro RC, Issel CJ. Detection of equine infectious anemia viral RNA in plasma samples from recently infected and long-term inapparent carrier animals by PCR.. J Clin Microbiol 1996 Jun;34(6):1481-7.
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