RESUMO
Hepatitis C virus (HCV) infection is a major cause of severe liver disease including liver cirrhosis and hepatocellular carcinoma. Genotyping became fundamental in the clinical assessment of patients with hepatitis C because the genotype of the hepatitis C virus determines the chance of therapeutic response and duration of treatment. We developed a new real-time PCR assay for genotyping of HCV with increased specificity due to a novel approach to dual-labeled probe design using oligodeoxyinosine linkers. The assay allows genotypes 1, 2, 3 to be distinguished and genotypes 4-6 with high specificity to be blocked. The analytical sensitivity (150 IU/ml) can be implemented. Of the 285 clinical samples genotyped using the developed assay, 45% were genotype 1; 6%, genotype 2; 49%, genotype 3. No discordant results with 5-UTR sequencing and commercial genotyping assays were obtained.
Assuntos
Primers do DNA/química , Hepacivirus/genética , Hepatite C/genética , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Primers do DNA/genética , Genótipo , Hepacivirus/química , Humanos , RNA Viral/químicaRESUMO
The study deals with the current topical problem in the development of a laboratory algorithm for the detection of early stages of HIV infection and for the creation of a monitoring system on this basis for the spread of drug-resistant HIV-1 strains. The paper presents the results of experimental examination of the most accessible health care methods for the differential diagnosis of early HIV infection, by using the test systems in Russia and gives practical recommendations on their application.
Assuntos
Western Blotting/métodos , Imunofluorescência/métodos , Infecções por HIV/diagnóstico , HIV-1/isolamento & purificação , Fármacos Anti-HIV/farmacologia , Diagnóstico Diferencial , Farmacorresistência Viral , Diagnóstico Precoce , Anticorpos Anti-HIV/sangue , Proteína gp41 do Envelope de HIV/imunologia , Infecções por HIV/sangue , HIV-1/efeitos dos fármacos , HIV-1/imunologia , Humanos , Sensibilidade e Especificidade , Ureia/química , Carga ViralRESUMO
A method for the production of Marburg virus in preparative amounts has been developed. It is based on sedimentation of the virus from blood plasma of infected guinea pigs by ultracentrifugation followed by purification in sucrose density gradient or gel chromatography on macroporous glass sorbents. The optimal terms of blood collection in infected animals were determined. Purified virus did not lose its biological activity. Concentrated virus preparations were studied by electron microscopy and polyacrylamide gel electrophoresis.
