title: Clinical utility of a quantitative polymerase chain reaction for diagnosis of cytomegalovirus disease in solid organ transplant patients.
abstract: BACKGROUND
  Accurate and rapid diagnosis of human cytomegalovirus (HCMV) disease in solid organ transplant patients remains a challenge. We evaluated the clinical utility of a quantitative polymerase chain reaction (QPCR) method to diagnose transplant patients with HCMV disease.
  
  
  METHODS
  A total of 429 plasma samples from 121 solid organ transplant patients were prospectively collected and evaluated for HCMV using a QPCR assay. To enhance the sensitivity of the QPCR assay, plasma samples were centrifuged in a manner designed to concentrate the virions before nucleic acid extraction. Quantitation was achieved by co-amplifying an internal quantitative standard (IS) that contained the same primer sequences as for HCMV. Polymerase chain reaction products were detected by hybridization to 96-well microtiter plates coated with either a HCMV- or an IS-specific probe.
  
  
  RESULTS
  A total of 103 patients had all samples negative by QPCR. None of the 103 patients developed HCMV disease during the study. In contrast, 18 patients showed at least 1 sample positive by the QPCR assay, but only 8 of these developed HCMV disease. The mean viral load value for patients without HCMV disease was 93 viral copies (vc) per ml of plasma (range: 35-325 vc/ml plasma) and for the 8 patients with HCMV disease was 67,686 vc/ml plasma (range: 167-1,325,000 vc/ml plasma) (P<0.001). Using a cut-off value of 100 vc/ml plasma and clinical diagnosis of HCMV disease, the QPCR assay showed a sensitivity of 100% and specificity of 99.1%.
  
  
  CONCLUSIONS
  HCMV viral load may be useful in the diagnosis of HCMV disease in solid organ transplant patients.
authors.affiliation: Department of Pathology, Virginia Commonwealth University, Richmond 23298-0248, USA. agonzalez@hsc.vcu.edu
authors.forename: A
authors.initials: A
authors.lastname: Ferreira-Gonzalez
authors.forename: R A
authors.initials: RA
authors.lastname: Fisher
authors.forename: L A
authors.initials: LA
authors.lastname: Weymouth
authors.forename: M R
authors.initials: MR
authors.lastname: Langley
authors.forename: L
authors.initials: L
authors.lastname: Wolfe
authors.forename: D S
authors.initials: DS
authors.lastname: Wilkinson
authors.forename: C T
authors.initials: CT
authors.lastname: Garrett
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D003587:Cytomegalovirus / Q000235:genetics*; D003586:Cytomegalovirus Infections / Q000097:blood / Q000175:diagnosis*; D004279:DNA, Viral / Q000032:analysis / Q000097:blood / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D016377:Organ Transplantation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral