title: Evaluation of diagnostic methods for the detection of cytomegalovirus in recipients of allogeneic stem cell transplants.
abstract: BACKGROUND
  Although several diagnostic methods are available for the surveillance of patients at risk of human cytomegalovirus (CMV) infection and disease, little data is available on their comparative performances in the diagnostic setting.
  
  
  OBJECTIVE
  To compare different assays for CMV detection, especially assays based on (quantitative) DNA and mRNA detection.
  
  
  METHODS
  Eight allogeneic bone marrow and stem cell transplant recipients at high risk for developing CMV disease (donor CMV-negative, recipient positive) were regularly tested for 7-20 weeks post-transplant by spin-amplification rapid culture from urine (viruria), antigenemia (pp65 assay), pp67 mRNA in whole blood (NASBA), and CMV DNA both qualitatively (in-house PCR, whole blood) and quantitatively (in-house PCR, plasma; Cobas Amplicor CMV Monitor Test, plasma and whole blood; Hybrid Capture, whole blood).
  
  
  RESULTS
  Four patients (50%) suffered CMV reactivation during follow-up. Out of 104 sample dates, 41 (39.4%) yielded a positive CMV result in at least one assay. Out of the 28 samples tested by all assays, the highest percentage of positive results was obtained with the in-house quantitative PCR (60.7%), followed by the Hybrid Capture system (39.3%), the Cobas Amplicor CMV Monitor Test, plasma version (35.7%), the Cobas Amplicor CMV Monitor Test, whole blood version (32.1%), in-house qualitative PCR (28.6%), and the mRNA assay (21.4%). Viruria was positive in one sample and pp65 antigenemia was found in two samples.
  
  
  CONCLUSIONS
  Despite a considerable incidence of CMV reactivations, pre-emptive anti-CMV chemotherapy prevented the development of CMV disease with the exception of one case. The molecular assays had superior sensitivity to conventional ones. The antigenemia assay proved unsuitable for the surveillance of hematological transplant patients. However, none of the tests recognized all timepoints with CMV reactivation. Further comparative studies are needed to determine their respective diagnostic values.
authors.affiliation: Institute for Medical Virology, Johann Wolfgang Goethe University Hospital, Frankfurt am Main 60054, Germany. w.preiser@em.uni-frankfurt.de
authors.forename: W
authors.initials: W
authors.lastname: Preiser
authors.forename: S
authors.initials: S
authors.lastname: Bräuninger
authors.forename: R
authors.initials: R
authors.lastname: Schwerdtfeger
authors.forename: U
authors.initials: U
authors.lastname: Ayliffe
authors.forename: J A
authors.initials: JA
authors.lastname: Garson
authors.forename: N S
authors.initials: NS
authors.lastname: Brink
authors.forename: S
authors.initials: S
authors.lastname: Franck
authors.forename: H W
authors.initials: HW
authors.lastname: Doerr
authors.forename: H F
authors.initials: HF
authors.lastname: Rabenau
mesh_terms: D000293:Adolescent; D000328:Adult; D000956:Antigens, Viral / Q000097:blood; D003587:Cytomegalovirus / Q000235:genetics*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000821:virology; D004279:DNA, Viral / Q000097:blood / Q000652:urine; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010750:Phosphoproteins / Q000097:blood; D016133:Polymerase Chain Reaction / Q000379:methods; D011933:Reagent Kits, Diagnostic; D014184:Transplantation, Homologous; D014763:Viral Matrix Proteins / Q000097:blood
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral; D010750:Phosphoproteins; D011933:Reagent Kits, Diagnostic; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa