RESUMO
The WHO approved protocol of pandemic influenza virus A/H1N1 identification by the method of one-step RT-PCR was adopted. The cost of research was decreased due to adoption of RNA purification method and utilizing of the common enzyme systems for RT-PCR.
Assuntos
Surtos de Doenças , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Diagnóstico Diferencial , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/epidemiologia , Mucosa Bucal/virologia , Sensibilidade e Especificidade , UcrâniaRESUMO
A diagnostic kit for detection of avian influenza virus by real-time polymerase chain reaction was developed. This kit allows to identify the influenza viruses type A and highly pathogenic strain of avian influenza virus H5N1. The diagnostic kit is universal and adopted for ABI PRISM SDS (Applied Biosystems), RotorGene (Corbett Research) and iQCycler (BioRad) PCR machines.
Assuntos
Virus da Influenza A Subtipo H5N1 , Influenza Aviária/virologia , Reação em Cadeia da Polimerase/métodos , Sequência de Aminoácidos , Animais , Aves , Virus da Influenza A Subtipo H5N1/genética , Virus da Influenza A Subtipo H5N1/isolamento & purificação , Virus da Influenza A Subtipo H5N1/patogenicidade , Dados de Sequência Molecular , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , VirulênciaRESUMO
The chemical synthesis of a modified gene fragment that codes IgG-binding domain of protein G of Streptococcus sp. (SPG) was done. Two copies of gene fragmentes SPG were cloned into plasmid vector pET-24(a), for expression of recombinant truncated protein G (rSPG) in the culture of Escherichia coli. IgG-binding capacity and specificity for rSPG was confirmed by model assay such interaction in vitro. Basing on the obtained data we affirm that truncated recombinant protein G is able to bind immunoglobulin G in mammalians and can be used for purification of IgG from sera by affinity chromatography or for synthesis of immunoassay conjugates with broad range specificity.
Assuntos
Proteínas de Bactérias , Técnicas Imunoenzimáticas/métodos , Imunoglobulina G/metabolismo , Proteínas Recombinantes , Streptococcus/metabolismo , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Streptococcus/genéticaRESUMO
Several Escherichia coli strains producing HIV-specific recombinant proteins including env-1, env-2, and gag sequences have been obtained using different gene engineering techniques. These proteins have been isolated and purified from bacterial lysates by ion exchange chromatography on DEAE-Toyopearl and Streamline-SP columns. Polyacrylamide gel electrophoresis and immunoblotting have been used to prove recombinant proteins purity and to identify them. These proteins have been shown to be adequate as antigen preparations for enzyme immunoassay test-systems aimed at anti-HIV antibodies detection.
Assuntos
HIV-1/metabolismo , HIV-2/metabolismo , Proteínas Virais/biossíntese , Cromatografia DEAE-Celulose , Eletroforese em Gel de Poliacrilamida , Escherichia coli/metabolismo , Engenharia Genética/métodos , Immunoblotting , Imunoquímica , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Virais/química , Proteínas Virais/isolamento & purificaçãoRESUMO
The results from comparative studies in the reactions of immunofluorescence, complement binding, diffusion precipitation, hemagglutination, solid-phase immunoenzyme analysis, histochemical variant of immunoenzyme analysis as tests for detection of cattle rotavirus in the process of its isolation from pathological material and adaptation to cell cultures are presented. The immunofluorescence reaction is shown to have an advantage over the other reactions.
