Sensitive non-isotopic DNA hybridisation assay or immediate-early antigen detection for rapid identification of human cytomegalovirus in urine.
A sensitive non-radioactive DNA hybridisation assay employing digoxigenin-labelled probes was compared with immediate-early antigen detection and conventional virus isolation for the identification of human cytomegalovirus (HCMV) in 249 urine samples. Of 44 specimens yielding HCMV by virus isolation, more were positive by DNA hybridisation (32; 73%) than by immediate-early antigen detection (25; 52%) (P = 0.05). The specificity of the hybridisation assay in 45 apparently falsely positive specimens was supported by detection of HCMV DNA in 40 of these specimens using the polymerase chain reaction. Many urine specimens may thus contain large amounts of non-viable virus or free viral DNA. Evaluation of various protocols for the extraction and denaturation of virus DNA prior to hybridisation showed that proteinase K digestion with phenol/chloroform extraction was the most sensitive and reliable procedure. We conclude that the non-radioactive DNA hybridisation assay described is a potentially valuable routine diagnostic test.
Kimpton
C P
CP
Department of Pathological Sciences, Medical School, University of Manchester, U.K.
Morris
D J
DJ
Corbitt
G
G
D000956:Antigens, Viral / Q000652:urine*; D001483:Base Sequence; D003587:Cytomegalovirus / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D015342:DNA Probes / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism / Q000652:urine*; D004076:Digoxigenin / Q000378:metabolism; D019286:Endopeptidase K; D006801:Humans; D017874:Immediate-Early Proteins; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases / Q000378:metabolism
D000956:Antigens, Viral; D015342:DNA Probes; D004279:DNA, Viral; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D004076:Digoxigenin