title: HCMV-DNA is detected more frequently than infectious virus in blood leucocytes of immunocompromised patients: a direct comparison of culture-immunofluorescence and PCR for detection of HCMV in clinical specimens.
abstract: In two studies comparing detection of human cytomegalovirus (HCMV) in 118 patients (93 of whom were immunocompromised) by the polymerase chain reaction (PCR) and virus isolation using either early antigen detection by culture-immunofluorescence or conventional cytopathic effect, DNA-PCR was found to be the most sensitive, followed by culture-immunofluorescence, then by cytopathic effect. Urine was inhibitory to the action of Taq polymerase; this was overcome by concentration of HCMV with PEG 6000 prior to gene amplification. Without PEG treatment, HCMV-DNA in 6 of the 11 specimens positive by culture-immunofluorescence was not detectable by PCR. In healthy seropositive individuals, HCMV-DNA was not detected in leucocytes. However, in immunocompromised patients with AIDS or transplants, and therefore at high risk of HCMV infection or reactivation, blood leucocytes were usually positive for HCMV-DNA (19/20), some for as long as 20 weeks after initial detection and persisting for long after culture-immunofluorescence became negative. Neither HCMV-RNA nor infectious HCMV were detected in the follow-up blood leucocyte specimens from immunocompromised patients who had detectable HCMV-DNA in these cells. These data suggest that persistence of HCMV-DNA in blood leucocytes of immunocompromised patients after reactivation or primary infection may be due to persistence of non-viable virus, residual HCMV genomic DNA, or latent HCMV-DNA.
authors.affiliation: Department of Virology, ICPMR, Westmead Hospital, NSW, Australia.
authors.forename: V M
authors.initials: VM
authors.lastname: Ratnamohan
authors.forename: J M
authors.initials: JM
authors.lastname: Mathÿs
authors.forename: A
authors.initials: A
authors.lastname: McKenzie
authors.forename: A L
authors.initials: AL
authors.lastname: Cunningham
mesh_terms: D001483:Base Sequence; D003587:Cytomegalovirus / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000276:immunology; D004279:DNA, Viral / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D016867:Immunocompromised Host; D007962:Leukocytes / Q000382:microbiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods
chemical_list: D004279:DNA, Viral