RESUMO
BACKGROUND: Lymphocryptovirus (LCV) is found in various non-human primates. As a herpesvirus naturally infecting gibbons it is closely related to human Epstein-Barr virus (EBV) with which it shares considerable genetic, biological and epidemiologic features. METHODS: We collected blood samples from 70 gibbons (51 Hylobates lar, 18 Hylobates pileatus and 1 Hylobates agilis) for further separation into serum and peripheral blood mononuclear cells (PBMC). RESULTS: Only 13 of 70 (18.6%) sera were serologically positive for human EBV IgG but 64 of 70 (91.4%) PBMCs yielded the partial LCV DNA polymerase gene by semi-nested PCR, which we subjected to direct sequencing. All sequences showed 84% nucleic acid and 91% amino acid identity to human EBV. Phylogenetic tree analysis demonstrated gibbon LCVs clustered separately from other gammaherpesvirinae but closely related to LCV of other species. CONCLUSIONS: Based on LCV DNA detection, we discovered a high prevalence of LCV infection among gibbons. Further characterization of non-human primate LCV might thus provide new insight into both evolution and pathogenicity of gammaherpesvirinae.
Assuntos
DNA Polimerase Dirigida por DNA/genética , Infecções por Herpesviridae/veterinária , Hylobates/virologia , Lymphocryptovirus/enzimologia , Lymphocryptovirus/genética , Infecções Tumorais por Vírus/veterinária , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/sangue , Sequência de Bases , DNA Viral/genética , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Infecções por Herpesviridae/sangue , Infecções por Herpesviridae/virologia , Hylobates/sangue , Lymphocryptovirus/isolamento & purificação , Masculino , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária , Alinhamento de Sequência , Análise de Sequência de DNA , Estudos Soroepidemiológicos , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/virologiaRESUMO
Influenza A virus subtype H5N1 causes a rapidly fatal systemic disease in domestic poultry and spreads directly from poultry to humans. The aim of this study was to develop a rapid, cost-saving and effective method for influenza A virus subtype H5N1 detection. The selected primer set was used in single-step RT-PCR for simultaneous detection in multiplex format of the 276-, 189-, and 131-bp fragments, corresponding to sequences specific for M, H5 and N1. The amplified DNA fragments were clearly separated by agarose gel electrophoresis. The sensitivity of this assay was about 10(3) copies/microL. Moreover, this method can be applied to detect not only avian but also human influenza A virus subtype H5N1. In conclusion, the highlights of this particular method are its rapidity and cost-effectiveness, thus rendering it feasible and attractive for large-scale screening at times of influenza A virus subtype H5N1 outbreak.
