title: Cytomegalovirus infection in organ-transplant recipients: diagnostic value of pp65 antigen test, qualitative polymerase chain reaction (PCR) and quantitative Taqman PCR.
abstract: BACKGROUND
  The human cytomegalovirus (CMV) is a major cause of morbidity and mortality in transplant patients. In this study, we compared the diagnostic value of pp65 antigen test, qualitative nested polymerase chain reaction (PCR), and quantitative Taqman PCR in predicting the clinical outcome of CMV infection.
  
  
  METHODS
  A total of 169 samples derived from 59 organ-transplant recipients (kidney n= 46, liver n= 11, kidney and pancreas n= 2) were analyzed. Peripheral blood leukocytes (PBL) were isolated using dextran gradient centrifugation, and 2 x 10 cells were analyzed for pp65 antigen by immunofluorescence. A crude DNA extract obtained from the same number of cells was used for qualitative nested PCR and quantitative Taqman PCR analysis. RESULTS.: The correlation coefficient of pp65 antigen test and Taqman PCR was R= 0.699 (P = 0.001). With cut-off values for pp65 antigen test set at greater than 10 positive nuclei per 2 x 10 PBL, sensitivity was 91%, and positive predictive value (PPV) was 70%. When the corresponding cut-off value for Taqman PCR was applied (>125000 genome copies per 2 x 10 PBL), a sensitivity of 83% and a PPV of 68% were found. Both assays allowed for the monitoring of successful antiviral therapy. Although qualitative nested PCR was highly sensitive (95%), it was less useful in predicting CMV disease (PPV 47%) and in therapy control.
  
  
  CONCLUSIONS
  Our data show that pp65 antigen test and Taqman PCR are almost equivalent in the monitoring of CMV infection and disease when identical cell numbers are used for both assays.
authors.affiliation: Abteilung Virologie, Institut für Medizinische Mikrobiologie and Hygiene, Universität Freiburg, Germany. mkoeng@ukl.uni-freiburg.de
authors.forename: Ursula
authors.initials: U
authors.lastname: Meyer-Koenig
authors.forename: Manfred
authors.initials: M
authors.lastname: Weidmann
authors.forename: Günter
authors.initials: G
authors.lastname: Kirste
authors.forename: Frank T
authors.initials: FT
authors.lastname: Hufert
mesh_terms: D000328:Adult; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000821:virology; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D016377:Organ Transplantation; D010750:Phosphoproteins / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D019914:Taq Polymerase; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000032:analysis*
chemical_list: D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D019914:Taq Polymerase