[Direct diagnostic methods for the demonstration of cytomegaloviruses in the blood].
CMV can be detected in blood with two techniques: isolation of CMV from the buffy-coat on human embryonic fibroblasts; CMV genome detection by DNA hybridization technique. Isolation of CMV from blood plated on embryonic fibroblasts necessitates a delay of 10-30 days. Using monoclonal antibodies we have developed a 96 hours test to diagnose viremia. Fibroblasts grown on coverslip in tubes are inoculated with buffy-coat. Indirect immunofluorescence test is performed using the monoclonal antibodies E 13, 48 and 96 hours post-infection. DNA-DNA hybridization technique necessitates purification of CMV DNA. CMV DNA is then labelled with 32 P by nick-translation. This labelled DNA is used as probe for detection of CMV genome in DNA extracted from leukocytes. The hybridization is virus-specific. Cytomegaloviremia correlates with active infection and is not indicate of the carrier state of blood donors. CMV genome research in blood can potentially identify infective donors. Further studies are needed to correlate CMV genome detection in blood with presence of CMV antibodies.
D001782:Blood Donors; D002353:Carrier State / Q000175:diagnosis; D003587:Cytomegalovirus / Q000032:analysis*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D004279:DNA, Viral / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006801:Humans; D009693:Nucleic Acid Hybridization; D014766:Viremia / Q000175:diagnosis*
D004279:DNA, Viral
Méthodes de diagnostic direct appliquées à la mise en évidence du cytomégalovirus dans le sang.