RESUMEN
The diagnostic oligonucleotide microarray for subtyping of human and animal influenza A viruses (IAVs) was developed. We proposed a simple method of the fluorescent labeling of genomic segments of all known IAVs subtypes, the composition of the hybridization buffer, as well as the software of the data processing. 48 IAVs strains of different subtypes were analyzed using our microarray. All of them were identified, while 45 of 48 strains were unambiguously subtyped.
Asunto(s)
Genoma Viral , Virus de la Influenza A/clasificación , Tipificación Molecular/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/instrumentación , Infecciones por Orthomyxoviridae/virología , ARN Viral/clasificación , Programas Informáticos , Animales , Humanos , Virus de la Influenza A/genética , Virus de la Influenza A/aislamiento & purificación , Gripe Humana/diagnóstico , Gripe Humana/virología , Dispositivos Laboratorio en un Chip , Infecciones por Orthomyxoviridae/diagnóstico , ARN Viral/genéticaRESUMEN
AIM: Development of technological stages of preparation of experimental influenza whole-virion inactivated adsorbed vaccine based on recombinant influenza virus strains NIBRG-14 and A/Astana/RG/6:2/2009. MATERIALS AND METHODS: 2 recombinant vaccines influenza strains were used in the study--NIBRG-14 and A/Astana/RG/6:2/2009. Purification of native virus-containing allantoic fluid was performed by ion-exchange chromatography. The virus was inactivated by formaldehyde. Merthiolate at concentration of 0.1 mg/ml was added to the vaccine as a preserving substance. Aluminium hydroxide was used as an adjuvant. Harmlessness and immunogenicity (HI) of the constructed preparation are determining. RESULTS: Virus-containing materials from recombinant strains with biological activity of 8.5 - 9.0 lg EID50/cm3 and hemagglutination activity of 1:256 - 1:1024 in chicken embryos were obtained. Optimal inactivation regimen of non-purified suspensions by formaldehyde was established and combined scheme of purification and concentration of influenza virus was selected that provide harmlessness and immunogenicity of experimental samples of inactivated vaccines against highly pathogenic influenza A/H5N1 in experiments in mice. CONCLUSION: The data obtained on quality parameters of intermediate products and final vaccine give evidence on their compliance with normative parameters for whole-virion influenza purified vaccine.
Asunto(s)
Anticuerpos Antivirales/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Gripe Humana/prevención & control , Infecciones por Orthomyxoviridae/prevención & control , Adyuvantes Inmunológicos , Hidróxido de Aluminio/inmunología , Animales , Anticuerpos Antivirales/sangre , Embrión de Pollo , Formaldehído/química , Hemaglutinación , Humanos , Inmunización , Vacunas contra la Influenza/biosíntesis , Gripe Humana/inmunología , Gripe Humana/virología , Kazajstán , Ratones , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/virología , Vacunas de Productos Inactivados , Vacunas Sintéticas , Virión/inmunología , Virión/aislamiento & purificaciónRESUMEN
Influenza reassortant viruses A/SPb/HK/09(H1N1), A/Astana/HK/2009 (H5N1), A/Otar/HK/2010(H3N8), and A/Perth/ HK/2011(H3N2), carrying surface antigens of different subtypes, were constructed on the basis of new potential unified donor strain A/HK/1/68/162/35(H3N2). The virulence and reproduction activity of the obtained reassortants were tested. The safety of the candidate live and inactivated influenza vaccines produced from the reassortant viruses was demonstrated. The study demonstrates that A/HK/1/68/162/35 can be used as a unified donor for attenuated and high-yield vaccine reassortants.