RESUMEN
The sensitivity and specificity of 4 experimental test systems for dot enzyme immunoassay (dot-EIA) and solid-phase lanthanide immunofluorescent analysis (SP LIFA) were studied with Venezuelan equine encephalomyelitis (VEE) and variolovaccinia viruses. Test systems for SP LIFA proved to be 25 times more sensitive than those for dot-EIA. The test systems were highly specific and did not react with the heterologous viruses and proteins. Diagnostic agent for detecting VEE virus in dot-EIA was false-positive with Sindbis virus in low dilutions. The first trials of the Russian test systems for dot-EIA and SP LIFA showed that these systems rapidly and reliably detect VEE and variolovaccinia viruses in liquid samples.
Asunto(s)
Virus de la Encefalitis Equina Venezolana/aislamiento & purificación , Técnica del Anticuerpo Fluorescente , Immunoblotting , Metales de Tierras Raras , Virus Vaccinia/aislamiento & purificación , Antígenos Virales/análisis , Virus de la Encefalitis Equina Venezolana/inmunología , Virus Vaccinia/inmunologíaRESUMEN
Human and animal sera were tested for the presence of antibodies to Venezuelan equine encephalomyelitis (VEE) virus by direct enzyme immunoassay (EIA). For the test, the plates were sensitized with a VEE virus preparation purified in a two-phase system of water-soluble polymers. The proposed EIA variant was as specific as that with VEE antigen obtained by fractionation on sucrose cushion, but more sensitive. The high specificity of the assay allowed the antigen purified in the water-polymer system to be used for investigation of antigen relationships among viruses of the VEE complex.
Asunto(s)
Anticuerpos Antivirales/sangre , Antígenos Virales/aislamiento & purificación , Virus de la Encefalitis Equina Venezolana/inmunología , Animales , Especificidad de Anticuerpos , Western Blotting , Electroforesis en Gel de Poliacrilamida , Virus de la Encefalitis Equina Venezolana/aislamiento & purificación , Humanos , Técnicas para Inmunoenzimas , Ratones , Conejos , Sensibilidad y Especificidad , Virología/métodosRESUMEN
Potentialities of differentiation between Venezuelan equine encephalomyelitis (VEE) complex viruses by time-resolved fluoroimmunoassay and enzyme immunoassay were studied. For this, 4 test systems were used based on different combinations of native and labeled polyclonal antibodies to VEE virus, strain Trinidad, and monoclonal (MCA) antibody MAK 14-7 to protein EL of this virus. The maximal sensitivity and specificity was achieved in the test system formed from native MCA MAK 14-7 for sensitization of the solid phase and labeled polyclonal immunoglobulins for demonstration of the test results. This combination of antibodies allowed to differentiate the epidemic variant of VEF/Trinidad (IA) from epizootic variants of Mucambo (III), Pixuna (IV) and attenuated strain No. 230.
Asunto(s)
Anticuerpos Monoclonales , Virus de la Encefalitis Equina Venezolana/clasificación , Metales de Tierras Raras , Animales , Anticuerpos Monoclonales/aislamiento & purificación , Anticuerpos Antivirales/inmunología , Anticuerpos Antivirales/aislamiento & purificación , Antígenos Virales/análisis , Antígenos Virales/inmunología , Antígenos Virales/aislamiento & purificación , Líquido Ascítico/inmunología , Virus de la Encefalitis Equina Venezolana/inmunología , Epítopos/análisis , Estudios de Evaluación como Asunto , Técnica del Anticuerpo Fluorescente , Técnicas para Inmunoenzimas , RatonesRESUMEN
Employment of radioimmunoassay led to the demonstration of serological crossing between tick-borne encephalitis (TBE) virus and Venezuelan equine encephalomyelitis (VEE) virus. Using hybridoma technology, three hybridomas were produced secreting monoclonal antibodies (MAb) cross-reacting with these two viruses. With MAb, the epitope of binding of these antibodies was shown to be located on protein E of TBE virus and protein E1 of VEE virus. Despite the low percentage (14%) of homology of amino acid sequences of these proteins, 12 areas with homology from 24% to 63% were demonstrated. Considering conservative replacements, homology of these areas was 53%-75%. The assumed existence of some of these areas in alpha-helical conformation may explain the observed immunological crossing.