Rapid detection of equine herpesvirus type-1 antigens in nasal swab specimens using an antigen capture enzyme-linked immunosorbent assay.
Abstract: An antigen capture enzyme-linked immunosorbent assay (ELISA) was developed for the detection of equine herpesvirus type-1 (EHV-1) antigens in nasal swab specimens. The test was designed as a solid phase, amplified sandwich assay in which an EHV-1 specific monoclonal antibody was used to capture virus antigen and polyclonal antisera used to detect antigen bound to the test plates. Eight monoclonal antibodies were tested for their ability to capture virus antigen and one was selected for routine use. The sensitivity and specificity of the ELISA was compared with that of virus isolation using swabs from ponies which were experimentally infected with EHV-1. Of 72 nasal swabs collected, 32 were found to be positive by both virus isolation (VI) and ELISA, a further 15 samples were positive by VI alone, but none of the samples were positive by ELISA and negative by VI. This yielded an overall assay sensitivity of 68% and specificity of 100%. The assay proved useful for diagnosis since virus antigen was detected during the first four days post-infection which corresponded to the acute phase of disease when some clinical symptoms were apparent. In addition, the assay could be completed within one day when antibody coated plates were available.
Publication Date: 1992-09-01 PubMed ID: 1331153DOI: 10.1016/0166-0934(92)90103-kGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Antibodies
- Antigen
- Antisera
- Clinical Study
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Equine Herpesvirus
- Horses
- Immunology
- Infection
- Infectious Disease
- Laboratory Methods
- Monoclonal Antibodies
- Pathology
- Veterinary Medicine
- Veterinary Research
- Virus
Summary
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The research involves the development of an antigen capture enzyme-linked immunosorbent assay (ELISA) for quick detection of equine herpesvirus type-1 (EHV-1) antigens present in nasal swab specimens. It compares the sensitivity and specificity of the test with virus isolation procedures on experimentally infected ponies.
Methodology and Results
- A solid-phase, amplified sandwich assay was engineered specifically for the detection of EHV-1 antigens. This assay design involves the use of a monoclonal antibody specifically attuned to EHV-1 to capture the virus antigen.
- Polyclonal antisera were then used to identify the antigen that had been captured and bonded to the test plates.
- A set of eight monoclonal antibodies were tested for their capability to capture virus antigen, one of which was selected for regular use based on its effectiveness.
- The researchers compared the sensitivity and specificity of the ELISA method with current standard virus isolation techniques. This comparison was made using nasal swab tests from ponies purposely infected with EHV-1.
- A total of 72 nasal swab samples were collected. Out of these, both virus isolation and ELISA identified 32 as positive. Fifteen more samples were found positive by virus isolation only, but no samples were identified as positive by ELISA and negative by virus isolation.
- From these results, the research team concluded that the ELISA demonstrated an overall sensitivity of 68% and a specificity of 100%.
Significance and Conclusion
- The assay is particularly useful for diagnosis as it was found to detect the virus antigen within the first four days post-infection. This timeline corresponds to the acute phase of the disease when some clinical symptoms are visible.
- Moreover, the research revealed that the ELISA process could be finished within a day if plates coated with antibodies were ready for use.
- The quick turn-around time and the high specificity of the ELISA assay offer advantages over the standard virus isolation technique, pointing to the potential of improving diagnostic methods for EHV-1.
Cite This Article
APA
Sinclair R, Mumford JA.
(1992).
Rapid detection of equine herpesvirus type-1 antigens in nasal swab specimens using an antigen capture enzyme-linked immunosorbent assay.
J Virol Methods, 39(3), 299-310.
https://doi.org/10.1016/0166-0934(92)90103-k Publication
Researcher Affiliations
- Department of Infectious Diseases, Animal Health Trust, Newmarket, Suffolk UK.
MeSH Terms
- Animals
- Antibodies, Monoclonal
- Antibodies, Viral
- Antigens, Viral / analysis
- Enzyme-Linked Immunosorbent Assay / methods
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / immunology
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horses
- Mice
- Mice, Inbred BALB C
- Nasal Cavity / microbiology
- Species Specificity
Citations
This article has been cited 1 times.- Singh BK, Yadav MP, Tewari SC. Neutralizing and complement-fixing monoclonal antibodies as an aid to the diagnosis of equine herpesvirus-1 infection. Vet Res Commun 2001 Dec;25(8):675-86.
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