RESUMO
A rapid, non-radioactive assay for the detection of proviral Simian immunodeficiency virus (SIV) in tissue-culture cells is described. The assay is based on the co-amplification of the SIV env and gag genes by the polymerase chain reaction (PCR). When the gag PCR product is blotted onto a nylon membrane and hybridised to a radioactive oligonucleotide probe, the assay can also be used to detect the SIV gag gene in DNA isolated directly from experimentally infected cynomolgus macaque lymphocytes. This provides a valuable assay for the presence of proviral SIV during animal trials of AIDS vaccines and chemotherapeutics.
Assuntos
Amplificação de Genes , Genes env , Genes gag , Reação em Cadeia da Polimerase , Infecções por Retroviridae/microbiologia , Vírus da Imunodeficiência Símia/isolamento & purificação , Animais , Sequência de Bases , Células Cultivadas , DNA Viral/análise , DNA Viral/genética , Linfócitos/microbiologia , Macaca fascicularis , Macaca mulatta , Dados de Sequência Molecular , Provírus/genética , Provírus/isolamento & purificação , Vírus da Imunodeficiência Símia/genéticaRESUMO
Negative staining electron microscopy was used to study sucrose gradient-purified preparations of the simian immunodeficiency virus (SIVmac251). Both isolated and aggregated virus particles were observed together with some free-lying virus cores. The cores were 110 nm long and 25 to 50 nm wide and were mainly conical or wedge-like in shape. Surface projections were seen on the envelope membrane of many of the virus particles; the knobs were approximately 6 nm in length, 10 nm wide and from an end-on view they had a Y or triangular-shaped morphology.
