RESUMEN
INTRODUCTION: Rotavirus infection is one of the main concerns in infectious pathology in humans, mammals and birds. Newborn piglets or rodents are usually being used as a laboratory model for the evaluation of immunogenicity and efficacy for all types of vaccines against rotavirus A (RVA), and the use of ELISA for the detection of virus-specific antibodies of specific isotype is an essential step of this evaluation. OBJECTIVE: Development of indirect solid-phase ELISA with VP2/VP6 rotavirus VLP as an antigen to detect and assess the distribution of RVA-specific IgG, IgM and IgA in the immune response to rotavirus A. MATERIALS AND METHODS: VP2/VP6 rotavirus VLP production and purification, electron microscopy, PAGE, immunoblotting, ELISA, virus neutralization assay. RESULTS: The study presents the results of development of a recombinant baculovirus with RVA genes VP2-eGFP/VP6, assessment of its infectious activity and using it for VLP production. The morphology of the VP2/VP6 rotavirus VLPs was assessed, the structural composition was determined, and the high antigenic activity of the VLP was established. VLP-based ELISA assay was developed and here we report results for RVA-specific antibody detection in sera of different animals. CONCLUSION: The developed ELISA based on VP2/VP6 rotavirus VLP as a universal antigen makes it possible to detect separately IgG, IgM and IgA antibodies to rotavirus A, outlining its scientific and practical importance for the evaluation of immunogenicity and efficacy of traditional vaccines against rotavirus A and those under development.
Asunto(s)
Rotavirus , Humanos , Recién Nacido , Animales , Porcinos , Rotavirus/genética , Proteínas Recombinantes , Anticuerpos Antivirales , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G , Inmunoglobulina A , Inmunidad , Inmunoglobulina M , Antígenos Virales/genética , MamíferosRESUMEN
Reverse genetics was applied to engineering of the reassortantvaccine candidate strain against highly pathogenic avian influenza viruses (HPAIVs) of the H5 subtype. The new strain recPR8-H5N1 contains the HA gene from the Russian HPAIV A/Kurgan/05/2005 (H5N1), the NA and internal genes from A/Puerto Rico/8/34 (H1N1). The strain recPR8-H5N1 demonstrated the antigenic specificity (H5), high proliferation rate in 12 days chicken embryos, and was lethal for the embryos in 36 hours. An inactivated emulsified vaccine based on the strain recPR8-H5N1 elicited high antibody titers and protected 6-week-old chickens from lethal challenge with the HPAIV A/Kurgan/05/2005 (H5N1) on day 21 after single immunization. Infection of non-vaccinated birds with the strain recPR8-H5N1 did not cause any pathology, and the virus was not detected using PCR in blood and cloacal swabs on day 7 p.i. Specific weak seroconversion caused by infection with the strain recPR8-H5N1 was detected on day 14 p.i. As a result, a new influenza virus strain was obtained with modified properties.
Asunto(s)
Anticuerpos Antivirales/biosíntesis , Antígenos Virales/genética , Subtipo H5N1 del Virus de la Influenza A/genética , Vacunas contra la Influenza/inmunología , Gripe Aviar/prevención & control , Virus Reordenados/genética , Genética Inversa , Animales , Antígenos Virales/inmunología , Embrión de Pollo , Pollos/virología , Ingeniería Genética/métodos , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Humanos , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H1N1 del Virus de la Influenza A/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/genética , Gripe Aviar/inmunología , Gripe Aviar/virología , Virus Reordenados/inmunología , Vacunas de Productos Inactivados , Replicación ViralRESUMEN
Results of monitoring of different subtypes of avian leukosis virus (ALV) from commercial poultry farms in 14 regions of Russian Federation were discussed. Only three regions were found to be negative. ALV was detected in other 11 regions in 46-64% cases (for different regions). The phylogenetic analysis of the genomes for the 12 field isolates of ALV was carried out in different regions of Russian Federation. The isolates belong to different subtypes of the virus and form two large groups. The genomic differences between Russian and foreign isolates within each group range from 5% to 10%.
