Development and validation of a monoclonal antibody blocking ELISA for the detection of antibodies against both equine herpesvirus type 1 (EHV1) and equine herpesvirus type 4 (EHV4).
Abstract: A monoclonal antibody blocking ELISA was developed for the detection of antibodies directed against either EHV1 or EHV4. For this purpose, we selected a monoclonal antibody directed against a cross-reactive, conservative and immunodominant epitope of both EHV1 and EHV4. High antibody titres were found in rabbit antisera and SPF-foal antisera infected with either EHV1 or EHV4. After experimental challenge of conventional horses with EHV1 or EHV4 significant increases in CF and ELISA titres were found, whereas VN antibodies did not always increase significantly. In 344 paired serum samples submitted for diagnostic purposes a good agreement (kappa = 0.75, confidence limits = 0.63-0.88) was found between VN test and ELISA regarding a significant increase in titres. Also, a good correlation was found between VN and ELISA titres (r = 0.76, p<<0.0005). The relative sensitivity and specificity of the Mab blocking ELISA as compared with the VN test were 99.9 and 71%, respectively. The rather low relative specificity of the ELISA may be explained by a relatively low sensitivity of the VN test. The ELISA also detected increases in titre after vaccination with an EHV1 subunit vaccine, and after primary field infections in weaned foals. We concluded that the Mab blocking ELISA is more sensitive, easier to perform, more rapid and more reproducible than the VN test. We consider this test as a valuable tool for serological diagnosis of both EHV1 and EHV4 infections.
Publication Date: 2000-02-09 PubMed ID: 10665532DOI: 10.1016/s0378-1135(99)00162-5Google Scholar: Lookup
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- Journal Article
Summary
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The research article presents the creation and verification of a monoclonal antibody blocking ELISA (Enzyme-Linked Immunosorbent Assay), which aids in detecting antibodies against equine herpesvirus type 1 (EHV1) and equine herpesvirus type 4 (EHV4).
Development of Monoclonal Antibody Blocking ELISA
- The study developed a monoclonal antibody blocking ELISA for the detection of antibodies targeted against EHV1 or EHV4.
- In order to achieve this, a monoclonal antibody aimed at a cross-reactive, conservative, and immunodominant epitope of both EHV1 and EHV4 was selected.
- High antibody titres were discovered in rabbit antisera and SPF-foal antisera infected with either EHV1 or EHV4.
Testing and Results
- Upon experimental challenge of conventional horses with EHV1 or EHV4, the researchers found significant increases in CF and ELISA titres, although VN antibodies did not always increase significantly.
- 344 paired serum samples were used for diagnostic purposes. Researchers found good agreement (kappa = 0.75, confidence limits = 0.63-0.88) between VN test and ELISA regarding a notable increase in titres.
- There was also a strong correlation between VN and ELISA titres (r = 0.76, p<<0.0005).
Relative Sensitivity and Specificity, and ELISA’s Advantage
- The relative sensitivity and specificity of the Mab blocking ELISA compared with the VN test was 99.9 and 71% respectively.
- The researchers noted that lower relative specificity of the ELISA could be due to the relatively low sensitivity of the VN test.
- The ELISA also detected increases in titre after vaccination with an EHV1 subunit vaccine and after primary field infections in weaned foals.
- The researchers concluded that the Mab blocking ELISA is more sensitive, easier to perform, quicker, and more reproducible than the VN test, making it a valuable tool for serological diagnosis of both EHV1 and EHV4 infections.
Cite This Article
APA
van Maanen C, de Boer-Luijtze E, Terpstra C.
(2000).
Development and validation of a monoclonal antibody blocking ELISA for the detection of antibodies against both equine herpesvirus type 1 (EHV1) and equine herpesvirus type 4 (EHV4).
Vet Microbiol, 71(1-2), 37-51.
https://doi.org/10.1016/s0378-1135(99)00162-5 Publication
Researcher Affiliations
- Animal Health Service, Deventer, The Netherlands. c.v.maanen@gdvdieren.nl
MeSH Terms
- Animals
- Antibodies, Monoclonal
- Antibodies, Viral / blood
- Complement Fixation Tests / veterinary
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / immunology
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / immunology
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horse Diseases / virology
- Horses
- Male
- Neutralization Tests / veterinary
- Rabbits
- Reproducibility of Results
- Sensitivity and Specificity
- Vaccination / veterinary
- Varicellovirus / immunology
- Varicellovirus / isolation & purification
Citations
This article has been cited 3 times.- Li X, Li G, Teng Q, Yu L, Wu X, Li Z. Development of a blocking ELISA for detection of serum neutralizing antibodies against newly emerged duck Tembusu virus. PLoS One 2012;7(12):e53026.
- Singh BK, Ahuja S, Gulati BR. Development of a neutralizing monoclonal antibody-based blocking ELISA for detection of equine herpesvirus 1 antibodies. Vet Res Commun 2004 Jul;28(5):437-46.
- Ali AAH, Abdallah F, Shemies OA, Kotb G, Nafea MR. Molecular characterization of equine herpes viruses type 1 and 4 among Arabian horse populations in Egypt during the period between 2021 and 2022. Open Vet J 2024 Jan;14(1):534-544.
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