Comparison of dot-blot DNA hybridisation and immediate early nuclear antigen production in cell culture for the rapid detection of human cytomegalovirus in urine.
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.
Morris
D J
DJ
Department of Medical Microbiology, Medical School, University of Manchester, U.K.
Lomax
J
J
Fox
A J
AJ
Corbitt
G
G
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
D000954:Antigens, Surface; D000956:Antigens, Viral; D004279:DNA, Viral; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus