title: A prospective study of a quantitative PCR ELISA assay for the diagnosis of CMV pneumonia in lung and heart-transplant recipients.
abstract: BACKGROUND
  Qualitative polymerase chain reaction (PCR) for the identification of cytomegalovirus (CMV) infection has a low predictive value for the identification of CMV pneumonia. This study prospectively evaluated the application of a quantitative PCR Enzyme-Linked Immuno-Sorbent Assay (ELISA) assay in 9 lung- and 18 heart-transplant recipients who did not receive ganciclovir prophylaxis.
  
  
  METHODS
  DNA was collected from peripheral blood polymorphonuclear leucocytes (PMNL) posttransplantation. Oligonucleotide primers for the glycoprotein B gene (149 bp) were used in a PCR ELISA assay using an internal standard for quantitation. CMV disease was defined as histological evidence of end organ damage.
  
  
  RESULTS
  The median level CMV genome equivalents in patients with CMV disease was 2665/2 x 10(5) PMNL (range 1,200 to 61,606) compared to 100 x 10(5) PMNL (range 20 to 855) with infection but no CMV disease (p = 0.036). All patients with CMV disease had genome equivalents levels of >1200/2 x 10(5) PMNL. A cut-off level of 1,200 PMNL had a positive predictive value for CMV disease of 100% and a negative predictive value of 100%. The first detection of levels of CMV genome equivalents above a level of 1200/2 x 10(5) PMNL was at a median of 58 days (range 47 to 147) posttransplant.
  
  
  CONCLUSIONS
  Quantitative PCR assays for the diagnosis of CMV infection may predict patients at risk of CMV disease and thereby direct preemptive treatment to high-risk patients.
authors.affiliation: Public Health Laboratory, Withington Hospital, Manchester, UK.
authors.forename: L
authors.initials: L
authors.lastname: Barber
authors.forename: J J
authors.initials: JJ
authors.lastname: Egan
authors.forename: J
authors.initials: J
authors.lastname: Lomax
authors.forename: Y
authors.initials: Y
authors.lastname: Haider
authors.forename: N
authors.initials: N
authors.lastname: Yonan
authors.forename: A A
authors.initials: AA
authors.lastname: Woodcock
authors.forename: A J
authors.initials: AJ
authors.lastname: Turner
authors.forename: A J
authors.initials: AJ
authors.lastname: Fox
mesh_terms: D000956:Antigens, Viral / Q000097:blood; D015331:Cohort Studies; D003587:Cytomegalovirus / Q000302:isolation & purification; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D004279:DNA, Viral / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016041:Heart-Lung Transplantation; D006801:Humans; D016040:Lung Transplantation; D009504:Neutrophils / Q000382:microbiology*; D011024:Pneumonia, Viral / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods; D011183:Postoperative Complications; D011237:Predictive Value of Tests; D011446:Prospective Studies
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral