ABSTRACT
Human erythropoietin (hEpo) production requires mammalian cells able to make complex post-translational modifications to guaranty its biological activity. As mammalian cell can be reservoir of pathogenic viruses and several animal origin components are usually used in the cultivation of mammalian cells, hEpo contamination with viruses is something of great concern. As consequence, this study investigated the viral removal and inactivation capacity of a recombinant-hEpo (rec-hEpo) purification process. Canine parvovirus, Human poliovirus type-2, Bovine viral diarrhea virus and Human immunodeficiency virus type-1 were used for measuring process viral removal and inactivation capacities. In conclusion, this study corroborated that the assessed rec-hEpo purification process has enough capacity (5.0-19.4 Logs) for removing and inactivating these model viruses and sodium hydroxide demonstrated to be a robust sanitization solution for chromatography columns (5.0 (PV-2)-6.7 (CPV) Logs).
Subject(s)
Disinfection/methods , Erythropoietin/isolation & purification , Virus Inactivation , Viruses/isolation & purification , Animals , CHO Cells , Cattle , Chromatography, Gel , Chromatography, Ion Exchange , Cricetinae , Cricetulus , Diarrhea Viruses, Bovine Viral/drug effects , Diarrhea Viruses, Bovine Viral/isolation & purification , Dogs , Erythrocyte Count , Erythropoietin/pharmacology , Female , HIV-1/drug effects , HIV-1/isolation & purification , HIV-1/ultrastructure , Humans , Kinetics , Mice , Microscopy, Electron, Transmission , Parvovirus, Canine/drug effects , Parvovirus, Canine/isolation & purification , Poliovirus/drug effects , Poliovirus/isolation & purification , Reproducibility of Results , Reticulocytes/cytology , Reticulocytes/drug effects , Sodium Hydroxide/pharmacology , Viruses/drug effectsABSTRACT
BACKGROUND: HIV-1 subtype B is largely predominant in the Caribbean, although other subtypes have been recently identified in Cuba. OBJECTIVES: To examine HIV-1 genetic diversity in Cuba. METHODS: The study enrolled 105 HIV-1-infected individuals, 93 of whom had acquired the infection in Cuba. DNA from peripheral blood mononuclear cells was used for polymerase chain reaction amplification and sequencing of pol (protease-reverse transcriptase) and env (V3 region) segments. Phylogenetic trees were constructed using the neighbour-joining method. Intersubtype recombination was analysed by bootscanning. RESULTS: Of the samples, 50 (48%) were of subtype B and 55 (52%) of diverse non-B subtypes and recombinant forms. Among non-B viruses, 12 were non-recombinant, belonging to six subtypes (C, D, F1, G, H and J), the most frequent of which was subtype G (n = 5). The remaining 43 (78%) non-B viruses were recombinant, with 14 different forms, the two most common of which were Dpol/Aenv (n = 21) and U(unknown)pol/Henv (n = 7), which grouped in respective monophyletic clusters. Twelve recombinant viruses were mosaics of different genetic forms circulating in Cuba. Overall, 21 genetic forms were identified, with all known HIV-1 group M subtypes present in Cuba, either as non-recombinant viruses or as segments of recombinant forms. Non-B subtype viruses were predominant among heterosexuals (72%) and B subtype viruses among homo- or bisexuals (63%). CONCLUSION: An extraordinarily high diversity of HIV-1 genetic forms, unparalleled in the Americas and comparable to that found in Central Africa, is present in Cuba.
Subject(s)
Genetic Variation , HIV Infections/virology , HIV-1/genetics , Cuba/epidemiology , Databases, Genetic , Female , Genes, pol , HIV Envelope Protein gp120/genetics , HIV Infections/epidemiology , HIV-1/classification , Humans , Male , Peptide Fragments/genetics , PhylogenyABSTRACT
Se estudiaron 6 muestras de heces fecales procedentes de igual número de niños hospitalizados que presentaban enfermedad diarréica aguda (EDA), y se efectuaron con las mismas las pruebas de Rotalex, PAGE e inoculación en cultivo celular, e inmunofluorescencia indirecta (IFI). Todas las muestras resultaron positivas por Rotalex y PAGE. En cuanto a cultivos, se realizaron 15 pases de las muestras en linea celular MA-104 y no se produjo efecto citopatogénico en ninguno de los casos. Se comprobó la presencia de rotavirus humanos en el interior de las células de los cultivos en 4 casos con el empleo de la técnica de IFI. Se produjo un suero utilizando la cepa SA-11 y se empleó el mismo con éxito en la IFI
Subject(s)
Culture Techniques , Fluorescent Antibody Technique , Rotavirus Infections/diagnosisABSTRACT
Se estudiaron 6 muestras de heces fecales procedentes de igual número de niños hospitalizados que presentaban enfermedad diarréica aguda (EDA), y se efectuaron con las mismas las pruebas de Rotalex, PAGE e inoculación en cultivo celular, e inmunofluorescencia indirecta (IFI). Todas las muestras resultaron positivas por Rotalex y PAGE. En cuanto a cultivos, se realizaron 15 pases de las muestras en linea celular MA-104 y no se produjo efecto citopatogénico en ninguno de los casos. Se comprobó la presencia de rotavirus humanos en el interior de las células de los cultivos en 4 casos con el empleo de la técnica de IFI. Se produjo un suero utilizando la cepa SA-11 y se empleó el mismo con éxito en la IFI
