ABSTRACT
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.
Subject(s)
Gene Amplification , Genes, env , Genes, gag , Polymerase Chain Reaction , Retroviridae Infections/microbiology , Simian Immunodeficiency Virus/isolation & purification , Animals , Base Sequence , Cells, Cultured , DNA, Viral/analysis , DNA, Viral/genetics , Lymphocytes/microbiology , Macaca fascicularis , Macaca mulatta , Molecular Sequence Data , Proviruses/genetics , Proviruses/isolation & purification , Simian Immunodeficiency Virus/geneticsABSTRACT
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.
