FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Veterinary microbiology1992; 33(1-4); 143-153; doi: 10.1016/0378-1135(92)90042-r

Diagnostic methods for African horsesickness virus using monoclonal antibodies to structural and non-structural proteins.

Abstract: A panel of 32 hybridoma cell lines secreting monoclonal antibodies (MAbs) reactive with African horsesickness virus serotype 4 (AHSV-4) has been developed. Four of the MAbs recognized the major core antigen VP7, twenty recognized the outer capsid protein VP2 and eight reacted with the non-structural protein NS1. With the VP7-specific MAbs a rapid and sensitive double antibody sandwich immunoassay has been developed to detect viral antigen in infected Vero cells and in spleen tissue from AHSV-infected horses. The sensitivity of the assay is 10 ng viral antigen per 100 microliters. The NS1-specific MAbs allowed visualization by immunofluorescence of tubule-like structures in the cytoplasm of infected Vero cells. This can be very useful as a confirmatory diagnostic procedure. The antigenic map of the outer capsid VP2 protein with MAbs is also reported.
Get Full Text
Publication Date: 1992-11-01 PubMed ID: 1481354DOI: 10.1016/0378-1135(92)90042-rGoogle 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.
LinkedInCopy
  • 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 research article involves the development of diagnostic methods for African horsesickness virus using monoclonal antibodies that react to the virus’ structural and non-structural proteins. The goal was to create a faster and more sensitive way to detect the disease in horses.

Development of Monoclonal Antibodies

  • The primary focus of this study was to create a panel of 32 hybridoma cell lines that secrete monoclonal antibodies (MAbs) that react specifically with African horsesickness virus serotype 4 (AHSV-4).
  • These 32 hybridoma cell lines were developed with various antibodies targeting different proteins of the virus. Four were specific to the major core antigen VP7, twenty recognized the outer capsid protein VP2, and eight reacted with the non-structural protein NS1.

Sandwich Immunoassay

  • Using the VP7-specific MAbs, a double antibody sandwich immunoassay was developed. The purpose of this assay is to detect the viral antigen in infected Vero cells and in spleen tissue from AHSV-infected horses.
  • The sandwich immunoassay has a high sensitivity level, being able to detect as little as 10 ng of viral antigen per 100 microliters. This indicates a high level of precision and reliability in identifying the virus at low concentration levels.

Visualizing Non-Structural Protein

  • The monoclonal antibodies specific to the non-structural protein NS1 were used to visualize tubule-like structures in the cytoplasm of infected Vero cells via immunofluorescence.
  • The ability to see these structures provides a beneficial tool in the diagnosis process, especially for confirmatory procedures where visual evidence of infection is important.

Mapping the Outer Capsid VP2 Protein

  • The study also reports on the antigenic map of the outer capsid VP2 protein with the MAbs. This mapping is crucial to understand the interactions between the virus and the antibodies, yielding key information to understand the mechanism of the disease and, possibly, insight into vaccine development strategies.

Cite This Article

APA
Ranz AI, Miguet JG, Anaya C, Venteo A, Cortés E, Vela C, Sanz A. (1992). Diagnostic methods for African horsesickness virus using monoclonal antibodies to structural and non-structural proteins. Vet Microbiol, 33(1-4), 143-153. https://doi.org/10.1016/0378-1135(92)90042-r

Publication

Veterinary microbiology
ISSN: 0378-1135
NlmUniqueID: 7705469
Country: Netherlands
Language: English
Volume: 33
Issue: 1-4
Pages: 143-153

Researcher Affiliations

Ranz, A I
  • Immunologia Y Genetica Aplicada S.A. (Ingenasa) Madrid, Spain.
Miguet, J G
    Anaya, C
      Venteo, A
        Cortés, E
          Vela, C
            Sanz, A

              MeSH Terms

              • African Horse Sickness / diagnosis
              • African Horse Sickness Virus / immunology
              • African Horse Sickness Virus / isolation & purification
              • Animals
              • Antibodies, Monoclonal / biosynthesis
              • Antibodies, Monoclonal / immunology
              • Antigens, Viral / immunology
              • Binding, Competitive
              • Blotting, Western
              • Capsid / immunology
              • Enzyme-Linked Immunosorbent Assay
              • Fluorescent Antibody Technique
              • Horses
              • Hybridomas
              • Mice
              • Neutralization Tests
              • Precipitin Tests
              • Vero Cells
              • Viral Proteins / immunology

              Citations

              This article has been cited 7 times.
              1. van Wyngaardt W, Mashau C, Wright I, Fehrsen J. Serotype- and serogroup-specific detection of African horsesickness virus using phage displayed chicken scFvs for indirect double antibody sandwich ELISAs. J Vet Sci 2013;14(1):95-8.
                doi: 10.4142/jvs.2013.14.1.95pubmed: 23388433google scholar: lookup
              2. Castillo-Olivares J, Calvo-Pinilla E, Casanova I, Bachanek-Bankowska K, Chiam R, Maan S, Nieto JM, Ortego J, Mertens PP. A modified vaccinia Ankara virus (MVA) vaccine expressing African horse sickness virus (AHSV) VP2 protects against AHSV challenge in an IFNAR -/- mouse model. PLoS One 2011 Jan 26;6(1):e16503.
                doi: 10.1371/journal.pone.0016503pubmed: 21298069google scholar: lookup
              3. Chiam R, Sharp E, Maan S, Rao S, Mertens P, Blacklaws B, Davis-Poynter N, Wood J, Castillo-Olivares J. Induction of antibody responses to African horse sickness virus (AHSV) in ponies after vaccination with recombinant modified vaccinia Ankara (MVA). PLoS One 2009 Jun 22;4(6):e5997.
                doi: 10.1371/journal.pone.0005997pubmed: 19543394google scholar: lookup
              4. Patel JR, Heldens JG. Immunoprophylaxis against important virus disease of horses, farm animals and birds. Vaccine 2009 Mar 13;27(12):1797-1810.
                doi: 10.1016/j.vaccine.2008.12.063pubmed: 19402200google scholar: lookup
              5. Heldens JG, Patel JR, Chanter N, Ten Thij GJ, Gravendijck M, Schijns VE, Langen A, Schetters TP. Veterinary vaccine development from an industrial perspective. Vet J 2008 Oct;178(1):7-20.
                doi: 10.1016/j.tvjl.2007.11.009pubmed: 18313956google scholar: lookup
              6. Martínez-Torrecuadrada JL, Díaz-Laviada M, Roy P, Sánchez C, Vela C, Sánchez-Vizcaíno JM, Casal JI. Serologic markers in early stages of African horse sickness virus infection. J Clin Microbiol 1997 Feb;35(2):531-5.
                doi: 10.1128/jcm.35.2.531-535.1997pubmed: 9003637google scholar: lookup
              7. Ma X, Zhang Y, Na L, Qi T, Ma W, Guo X, Wang XF, Wang X. Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against African Horse Sickness Virus Serotype 1 VP2 Protein. Viruses 2024 Nov 15;16(11).
                doi: 10.3390/v16111780pubmed: 39599893google scholar: lookup
              Subscribe to our newsletter
              Join 99,970 horse owners like you who receive our email updates on horse health and nutrition.