Detection of cytomegaloviremia using monoclonal antibodies.
Isolation of cytomegalovirus (CMV) from blood plated on embryonic fibroblasts necessitates a delay of 10-30 days. Using monoclonal antibodies (McAb) we have developed a 96 hour test to diagnose viremia. We produced monoclonal antibodies directed against CMV (strain AD 169). Clone E 13 McAb detected a CMV early antigen in nuclei using indirect immunofluorescence. These monoclonal antibodies recognized reference strains (Davis and Towne) and 20 field isolates of CMV. E 13 McAb were therefore selected for the diagnostic tests. MRC 5 cells grown on coverslips in tubes were inoculated with buffy-coat from 51 blood samples from bone marrow allograft recipients and Guillain Barre polyneuritis, and centrifuged on hour at 4000 g at 36 degrees C. The indirect immunofluorescence test (IIF) was performed using E 13 McAb, 48 and 96 hours post infection. Three viremia were detected within 48 hours and five others within 96 hours. Plasma and buffy-coat were also inoculated onto MRC 5 cells grown in flasks. Flasks were observed daily for CMV cytopathic effect (CPE). CPE appeared within an average of 14 days. When CMV was detected by CPE, the IF tests were always positive within 96 hours after drawing the blood.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D003587:Cytomegalovirus / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000276:immunology; D003588:Cytopathogenic Effect, Viral; D005455:Fluorescent Antibody Technique; D006801:Humans; D014766:Viremia / Q000175:diagnosis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral