Laboratory diagnosis of African horse sickness: comparison of serological techniques and evaluation of storage methods of samples for virus isolation.
Abstract: Five serological methods of diagnosing African horse sickness were evaluated, using a battery of serum samples from experimental horses vaccinated and challenged with each serotype of African horse sickness virus (AHSV1 through AHSV9): agar gel immunodiffusion (AGID), indirect fluorescent antibody (IFA), complement fixation (CF), virus neutralization (VN), and enzyme-linked immunosorbent assay (ELISA). The 5 tests were also compared using a panel of field samples, convalescent equine sera with antibodies to domestic equine viral diseases, and sera from horses awaiting export. The ELISA described in this paper was group specific. It did not require calibration with a standard positive serum but did yield elevated values with negative sera that were repeatedly frozen and thawed or heat inactivated. The IFA test was sensitive but could not be used on some field sera as the control cells exhibited fluorescence, possibly due to the animal being recently vaccinated with cell culture material. Sixty-two experimental sera were compared by VN, CF, AGID, and ELISA. Forty sera, 10 positive and 30 negative, were correctly classified by the 5 serologic assays. The 22 remaining sera gave mixed reactions. The AGID had no false positive results but had false negative results for up to 20% of the samples, depending upon the comparison. The VN, CF, and ELISA were similar in their variability. The length of time that virus could be recovered from a viremic blood sample was compared in an evaluation of storage methods for virus isolation samples. Washed erythrocytes were held at 4 C, washed erythrocytes plus stabilizer were held at -70 C, and blood that was drawn into a preservative (oxalate/phenol/glycerol) was held at 4 C.(ABSTRACT TRUNCATED AT 250 WORDS)
Publication Date: 1990-01-01 PubMed ID: 2128615DOI: 10.1177/104063879000200108Google Scholar: Lookup
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- Comparative Study
- Journal Article
- African Horse Sickness
- Antibodies
- Complement Fixation
- Diagnosis
- Diagnostic Technique
- Disease
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Horses
- Immunofluorescence Assay
- Immunology
- Laboratory Methods
- Serodiagnosis
- Serological Surveys
- Serotypes
- Veterinary Medicine
- Veterinary Research
- Virology
- Virus
Summary
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The research article focuses on the evaluation of various serological techniques for diagnosing African horse sickness and analyses the effectiveness of different methods for collecting and storing samples for virus isolation.
Comparison of Serological Techniques
- The article starts with an investigation into five serological methods – agar gel immunodiffusion (AGID), indirect fluorescent antibody (IFA), complement fixation (CF), virus neutralization (VN), and enzyme-linked immunosorbent assay (ELISA) to identify the most effective way to diagnose African horse sickness.
- Experiments were conducted using serum samples from horses that had been vaccinated and challenged with different serotypes of the African horse sickness virus.
- Each of these methods was compared under varying conditions involving samples from the field, horses recovering from local viral diseases, and horses awaiting export.
Evaluation of ELISA and IFA Tests
- Secondary analysis of the ELISA test showed that it was group-specific and didn’t require calibration with a standard positive serum. However, it presented higher values with negative sera that were repeatedly frozen, thawed, or heat inactivated.
- The IFA test was noted to be sensitive but demonstrated limitations as it couldn’t be used on some field sera. This was due to an observed fluorescence in control cells which could have been caused by a recent vaccination with cell culture material.
Evaluation of VN, CF, AGID, and ELISA Tests
- Further, forty serum samples, including ten positive and thirty negative, were tested using VN, CF, AGID, and ELISA methods. The tests correctly classified all of the samples.
- However, 22 other samples demonstrated mixed reactions with these tests, indicative of variability in results with certain types of serum.
- Overall, AGID demonstrated no false-positive results but had up to 20% false negatives. The other three tests showed similar variability.
Comparison of Sample Storage Methods
- Lastly, the study evaluated how long a blood sample remained viable for virus recovery, considering three different sample storage methods.
- The storage methods included washing and preserving erythrocytes at 4 degrees Celsius, storing washed erythrocytes with a stabilizer at -70 degrees Celsius, and storing oxalate/phenol/glycerol preserved blood at 4 degrees Celsius.
Cite This Article
APA
House C, Mikiciuk PE, Berninger ML.
(1990).
Laboratory diagnosis of African horse sickness: comparison of serological techniques and evaluation of storage methods of samples for virus isolation.
J Vet Diagn Invest, 2(1), 44-50.
https://doi.org/10.1177/104063879000200108 Publication
Researcher Affiliations
- Foreign Animal Disease Diagnostic Laboratory, USDA, APHIS, Greenport, NY 11944.
MeSH Terms
- African Horse Sickness / diagnosis
- African Horse Sickness Virus / immunology
- African Horse Sickness Virus / isolation & purification
- Animals
- Antibodies, Viral / blood
- Blood Preservation
- Complement Fixation Tests
- Enzyme-Linked Immunosorbent Assay
- Fluorescent Antibody Technique
- Horses
- Immunodiffusion
- Neutralization Tests
Citations
This article has been cited 6 times.- Penzhorn L, Crafford JE, Guthrie AJ. Enhancing African horse sickness virus detection: comparing and adapting PCR assays. J Vet Diagn Invest 2026 Feb 7;:10406387261417355.
- Fairbanks EL, Brennan ML, Mertens PPC, Tildesley MJ, Daly JM. Re-parameterization of a mathematical model of African horse sickness virus using data from a systematic literature search. Transbound Emerg Dis 2022 Jul;69(4):e671-e681.
- Feldmann F, Shupert WL, Haddock E, Twardoski B, Feldmann H. Gamma Irradiation as an Effective Method for Inactivation of Emerging Viral Pathogens. Am J Trop Med Hyg 2019 May;100(5):1275-1277.
- Dennis SJ, Meyers AE, Guthrie AJ, Hitzeroth II, Rybicki EP. Immunogenicity of plant-produced African horse sickness virus-like particles: implications for a novel vaccine. Plant Biotechnol J 2018 Feb;16(2):442-450.
- Bachanek-Bankowska K, Maan S, Castillo-Olivares J, Manning NM, Maan NS, Potgieter AC, Di Nardo A, Sutton G, Batten C, Mertens PP. Real time RT-PCR assays for detection and typing of African horse sickness virus. PLoS One 2014;9(4):e93758.
- Venter M, Human S, Zaayman D, Gerdes GH, Williams J, Steyl J, Leman PA, Paweska JT, Setzkorn H, Rous G, Murray S, Parker R, Donnellan C, Swanepoel R. Lineage 2 west nile virus as cause of fatal neurologic disease in horses, South Africa. Emerg Infect Dis 2009 Jun;15(6):877-84.
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