title: Comparison of dot-blot DNA hybridisation and immediate early nuclear antigen production in cell culture for the rapid detection of human cytomegalovirus in urine.
abstract: The sensitivity and specificity of four modes of two assays, immediate early nuclear antigen detection in cell culture (IENAD) at 24 and 48 h post-infection (p.i.) by immunofluorescence using a murine monoclonal antibody, and dot-blot DNA hybridisation with overnight or prolonged autoradiography using the 32P-labelled HindIII J fragment of human cytomegalovirus (HCMV) DNA as probe, were compared for the rapid detection of HCMV in urine. The sensitivity of IENAD was enhanced by low-speed centrifugation at the time of inoculation. DNA hybridisation with overnight autoradiography was significantly less sensitive than IENAD at 24 h p.i. (P less than 0.001), and even with prolonged autoradiography the hybridisation assay was slower and significantly less sensitive than IENAD at 48 h p.i. (P less than 0.02). The specificity of the two assays was virtually 100%. The sensitivity of DNA hybridisation was thus clearly inferior to that of IENAD.
authors.affiliation: Department of Medical Microbiology, Medical School, University of Manchester, U.K.
authors.forename: D J
authors.initials: DJ
authors.lastname: Morris
authors.forename: J
authors.initials: J
authors.lastname: Lomax
authors.forename: A J
authors.initials: AJ
authors.lastname: Fox
authors.forename: G
authors.initials: G
authors.lastname: Corbitt
mesh_terms: D000954:Antigens, Surface / Q000235:genetics*; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics*; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000652:urine*; D004279:DNA, Viral / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins; D009693:Nucleic Acid Hybridization
chemical_list: D000954:Antigens, Surface; D000956:Antigens, Viral; D004279:DNA, Viral; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus