title: Improved monitoring of cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation by an ultrasensitive plasma DNA PCR assay.
abstract: Cytomegalovirus (CMV) DNA amplification assays in plasma have shown limited sensitivity compared to the detection of pp65 antigen in leukocytes. Our goal was to increase the sensitivity of a commercial CMV DNA PCR quantitative assay. After modification, the new assay was able to reproducibly detect 20 CMV DNA copies/ml of plasma. We compared this new ultrasensitive PCR assay with the standard PCR and the pp65 test for CMV detection and quantification in 22 consecutive allogeneic hematopoietic stem cell recipients. CMV infection or reactivation was detected in 84 of 319 (26%) samples by the ultrasensitive PCR assay compared to 38 of 319 (12%) samples by the pp65 assay (P < 0.01). All samples positive by the pp65 assay were positive by the ultrasensitive PCR, and CMV episodes were detected on average 4 days earlier and 7 days later than the first and the last pp65-positive test, respectively. In addition, during CMV episodes, the ultrasensitive assay identified positive samples that were inconsistently detected by the pp65 assay. The ultrasensitive assay was also much more sensitive than the standard PCR, with 26 versus 12% of CMV DNA-positive samples (P < 0.01). This assay improved the monitoring of CMV infection or reactivation in hematopoietic allogeneic stem cell recipients.
authors.affiliation: Division of Infectious Diseases. Division of Hematology. Division of Oncology, University Hospital of Geneva, Geneva, Switzerland. laurent.kaiser@hcuge.ch
authors.forename: Laurent
authors.initials: L
authors.lastname: Kaiser
authors.forename: Luc
authors.initials: L
authors.lastname: Perrin
authors.forename: Bernard
authors.initials: B
authors.lastname: Chapuis
authors.forename: Karine
authors.initials: K
authors.lastname: Hadaya
authors.forename: Lenka
authors.initials: L
authors.lastname: Kolarova
authors.forename: Christelle
authors.initials: C
authors.lastname: Deffernez
authors.forename: Saadia
authors.initials: S
authors.lastname: Huguet
authors.forename: Claudine
authors.initials: C
authors.lastname: Helg
authors.forename: Werner
authors.initials: W
authors.lastname: Wunderli
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D003587:Cytomegalovirus / Q000302:isolation & purification* / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000821:virology*; D004279:DNA, Viral / Q000097:blood*; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010750:Phosphoproteins / Q000097:blood; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D014184:Transplantation, Homologous / Q000009:adverse effects*; D014763:Viral Matrix Proteins / Q000097:blood
chemical_list: D004279:DNA, Viral; D010750:Phosphoproteins; D011933:Reagent Kits, Diagnostic; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa