RESUMEN
Two different methods were used to prepare subunit components of transmissible gastroenteritis (TGE) virus. The ultrastructure of the peplomer glycoprotein E2 of TGE virus was studied. The presence of detergent NP-40 was shown to prevent the formation of this glycoprotein aggregates. In dialysed preparations, the peplomers are aggregated to form multimer structures, the so-called "rosettes". The orientation of the peplomers in the "rosettes" and in the whole virion was identical. The antigenic activity of viral subunits was studied in guinea pigs. Preparations of isolated peplomers were shown to induce enhanced production of virus-neutralizing antibodies whose mean titre exceeded that in the group of animals immunized with whole virion-based preparations.
Asunto(s)
Antígenos Virales/inmunología , Coronaviridae/inmunología , Gastroenteritis Porcina Transmisible/inmunología , Virus de la Gastroenteritis Transmisible/inmunología , Animales , Anticuerpos Antivirales/inmunología , Formación de Anticuerpos , Antígenos Virales/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida , Técnica del Anticuerpo Fluorescente , Gastroenteritis Porcina Transmisible/prevención & control , Glicoproteínas/aislamiento & purificación , Glicoproteínas/ultraestructura , Cobayas , Inmunización , Microscopía Electrónica , Formación de Roseta , PorcinosRESUMEN
Two variants of enzyme immunoassay (EIA) using purified swine transmissible gastroenteritis virus antigen for sensitization of the solid phase and using infected cells on the solid phase have been developed for quantitation of specific antibodies to swine transmissible gastroenteritis (STG) virus. Both variants of the method were found to have approximately similar sensitivity and to be highly specific. The developed EIA proved to be approximately 100 times as sensitive as neutralization test. The optimal concentration of purified antigen for sensitization of microplates, as well as the multiplicity of infection of the heterologous cell culture grown in microplate wells have been determined. The above methods may be widely used in virological and epizootiological studies.
Asunto(s)
Anticuerpos Antivirales/análisis , Coronaviridae/inmunología , Virus de la Gastroenteritis Transmisible/inmunología , Animales , Especificidad de Anticuerpos , Células Cultivadas , Estudios de Evaluación como Asunto , Gastroenteritis Porcina Transmisible/diagnóstico , Técnicas para Inmunoenzimas/instrumentación , Pruebas de Neutralización , Porcinos , Virus de la Gastroenteritis Transmisible/aislamiento & purificaciónRESUMEN
Direct and indirect variants of enzyme immunoassay (EIA) were developed for the detection of specific antigen of swine transmissible gastroenteritis virus (TGEV). Preparation of IgG were obtained from highly specific hyperimmune sera of guinea pigs and gnotobiotic piglets and their optimal concentration for the tests were determined. For the direct EIA, a peroxidase conjugate was prepared from guinea pig IgG and its working dilution was determined. The developed EIA systems proved to be highly sensitive for the detection of TGEV antigen both in infected cell cultures and clinical specimens from sick piglets detecting 5 ng of the virus-specific protein.
Asunto(s)
Antígenos Virales/análisis , Coronaviridae/inmunología , Virus de la Gastroenteritis Transmisible/inmunología , Animales , Estudios de Evaluación como Asunto , Vida Libre de Gérmenes , Cobayas , Sueros Inmunes/aislamiento & purificación , Inmunización , Técnicas para Inmunoenzimas , Inmunoglobulina G/aislamiento & purificación , PorcinosRESUMEN
Pretreatment of the transmissible gastroenteritis virus of swine (TGE) (coronavirus) with trypsin increased the yield of the infectious virus approximately 100-fold when propagated in roller cultures of SPEV cells, the time required for the development of marked CPE being twice shorter than without treatment. The method of concentration and purification of TGE virus is described which yields a preparative amount of highly purified virions retaining their structure. Such preparations were used for immunization of guinea pigs. It is concluded that purified preparations of TGE virus possess marked antigenic potency and may be used for generation of highly active and specific hyperimmune sera.