RESUMEN
The article substantiates possibility of application of point enzyme-linked immunosorbent assay (dot-technique) for detecting viral antigens in samples from patients. To diagnose adenovirus infection conjugate of virus-specific monoclonal antibodies and peroxidase of horse-radish were used The chromatographic rectification of conjugate from free peroxidase permits diminishing background coloring of nitrocellulose membrane and therefore to increase sensitivity. The application of direct conjugates on the basis of virus-specific monoclonal antibodies increases specifcity of dot-technique and significantly shortens time period of analysis. As in case of application of direct conjugates on the basis of polyclonal serum, samples from patients require preliminary processing with detergent for preventing non-specific reactions. The dot-technique demonstrates good coincidence with data of polymerase chain reaction and after clinical trials it can be used in diagnostic of human viral infections.
Asunto(s)
Infecciones por Adenoviridae/diagnóstico , Adenoviridae/aislamiento & purificación , Anticuerpos Monoclonales/química , Anticuerpos Antivirales/química , Antígenos Virales/sangre , Immunoblotting/métodos , Adenoviridae/química , Adenoviridae/inmunología , Infecciones por Adenoviridae/inmunología , Infecciones por Adenoviridae/virología , Especificidad de Anticuerpos , Pruebas de Inhibición de Hemaglutinación , Peroxidasa de Rábano Silvestre/química , Humanos , Inmunoconjugados/química , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Group B streptococcal (GBS) gene encoding the putative lipoprotein and adherence factor ScaAB was cloned and expressed in E. coli. Recombinant ScaAB protein was isolated. Signal sequence of ScaAB was found to be cleaved in the E. coli host. ScaAB recombinant protein was immunogenic in mice and antibodies against this protein were discovered in mice sera after GBS infection. The perspectives of the use of ScaAB protein in GBS vaccine are discussed.
Asunto(s)
Proteínas Bacterianas/inmunología , Proteínas de la Membrana/inmunología , Proteínas Recombinantes/inmunología , Infecciones Estreptocócicas/prevención & control , Streptococcus agalactiae/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Antibacterianos/sangre , Adhesión Bacteriana , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Inmunización , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Datos de Secuencia Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Infecciones Estreptocócicas/microbiología , Infecciones Estreptocócicas/mortalidad , Streptococcus agalactiae/aislamiento & purificación , Streptococcus agalactiae/patogenicidadRESUMEN
The cold-adapted temperature-sensitive (ts) influenza virus strain A/Leningrad/134/17/57 (H2N2) multiplied well at 32 degrees C (optimal temperature); lower titres of infectious virus were obtained in developing chick embryos at 40 degrees C. In a canine kidney (MDCK) cell line and in primary calf kidney (CK) cells an increased reproduction of the virus was found at 40 degrees C especially in the presence of trypsin. The ratios of virus titres obtained at optimal versus higher temperatures (RCT40) were by 1,000 times lower than those found in chick embryos. Polyacrylamide gel electrophoresis revealed a comparable synthesis of the cold-adapted influenza virus strain polypeptides HA, NP, M and NS in MDCK cells, regardless whether they were incubated at optimal or non-permissive temperatures.