Diagnostic methods for African horsesickness virus using monoclonal antibodies to structural and non-structural proteins.
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.
Ranz
A I
AI
Immunologia Y Genetica Aplicada S.A. (Ingenasa) Madrid, Spain.
Miguet
J G
JG
Anaya
C
C
Venteo
A
A
Cortés
E
E
Vela
C
C
Sanz
A
A
D000355:African Horse Sickness / Q000175:diagnosis*; D000356:African Horse Sickness Virus / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D001667:Binding, Competitive; D015153:Blotting, Western; D002213:Capsid / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006736:Horses; D006825:Hybridomas; D051379:Mice; D009500:Neutralization Tests; D011233:Precipitin Tests; D014709:Vero Cells; D014764:Viral Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D014764:Viral Proteins