title: Novel DNA assay for cytomegalovirus detection: comparison with conventional culture and pp65 antigenemia assay.
abstract: We compared conventional cytomegalovirus (CMV) isolation, rapid viral culture, a CMV pp65 antigenemia assay, and a novel CMV DNA hybrid capture system (HCS). A total of 309 blood samples from individuals in different risk groups were assessed by at least two of the methods mentioned above. Leukocytes were recovered either after centrifugation in Leucosep tubes containing 1.080 Ficoll for pp65 assay or after simple differential lysis steps for DNA detection. HCS was based on DNA hybridization with a CMV RNA probe and its capture by antibodies to DNA-RNA hybrids. The CMV pp65 lower matrix protein was detected by fluorescence with c10-c11 monoclonal antibody in formalin-fixed leukocytes. Concordant results were observed for 92.9, 78.3, and 82.7% of the patients when comparing (i) viral culture and the pp65 antigenemia assay, (ii) viral culture and HCS, and (iii) the pp65 antigenemia assay and HCS, respectively. Discordant results were observed between a positive HCS result and negative culture and/or pp65 results. These results were associated with relatively low DNA levels (< 20 pg/10(6) cells) and positive viruria. In conclusion, the pp65 antigenemia assay is a rapid and reliable method of detecting CMV and is preferable to culture, but the Murex HCS appears to be more sensitive for CMV detection.
authors.affiliation: Laboratoire de Virologie, Centre Hospitalier Universitaire Angers, France.
authors.forename: N
authors.initials: N
authors.lastname: Veal
authors.forename: C
authors.initials: C
authors.lastname: Payan
authors.forename: D
authors.initials: D
authors.lastname: Fray
authors.forename: L
authors.initials: L
authors.lastname: Sarol
authors.forename: O
authors.initials: O
authors.lastname: Blanchet
authors.forename: S
authors.initials: S
authors.lastname: Kouyoumdjian
authors.forename: F
authors.initials: F
authors.lastname: Lunel
mesh_terms: D000956:Antigens, Viral / Q000097:blood; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000821:virology; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics*; D003951:Diagnostic Errors; D005069:Evaluation Studies as Topic; D006801:Humans; D009693:Nucleic Acid Hybridization; D010750:Phosphoproteins / Q000097:blood / Q000276:immunology; D012680:Sensitivity and Specificity; D014763:Viral Matrix Proteins / Q000097:blood / Q000276:immunology; D014773:Virology / Q000379:methods* / Q000706:statistics & numerical data; D014776:Virus Cultivation / Q000379:methods / Q000706:statistics & numerical data
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa