title: Comparison of LightCycler-based PCR, COBAS amplicor CMV monitor, and pp65 antigenemia assays for quantitative measurement of cytomegalovirus viral load in peripheral blood specimens from patients after solid organ transplantation.
abstract: In order to evaluate the LightCycler-based PCR (LC-PCR) as a diagnostic assay technique, a classical pp65 antigenemia assay and the commercially available COBAS Amplicor CMV Monitor (CACM) assay were compared to the LC-PCR assay for the detection and quantitation of cytomegalovirus (CMV) load in 404 parallel specimens of peripheral blood from 66 patients after solid organ transplantation. A good correlation existed among these three assays (r congruent with 0.6, P < 0.0001). The LC-PCR assay was the most sensitive (54% of specimens positive) compared to the CACM (48.6%) and the pp65 antigenemia (26%) assays. The LC-PCR assay detected all samples found positive by using both the CMV pp65 antigenemia assay and the CACM assay. The LC-PCR also had the widest dynamic range (from 250 to 10(7) DNA copies/ml of plasma). No cross-reactions were found among CMV and Epstein-Barr virus, varicella-zoster virus, or herpes simplex virus in the LC-PCR by using amplification with specifically designed primer pairs. Precision, expressed as the coefficient of variation, was <3% with standard DNA from cell cultures and between 6.55 and 14.1% with clinical specimens in repeat LC-PCR runs. One run of the LC-PCR took half of the time required for the semiautomated CACM procedure. Because of its sensitivity, specificity, cost-effectiveness, and simplicity, the LC-PCR assay could replace the pp65 antigenemia and the CACM assays as the preferred technique for the surveillance, diagnosis, and monitoring of response of CMV diseases in high-risk populations.
authors.affiliation: Provincial Laboratory for Public Health (Microbiology), University of Alberta Hospital, University of Alberta, Edmonton, Alberta T6G 2J2, Canada.
authors.forename: Xiaoli L
authors.initials: XL
authors.lastname: Pang
authors.forename: Linda
authors.initials: L
authors.lastname: Chui
authors.forename: Jayne
authors.initials: J
authors.lastname: Fenton
authors.forename: Barbara
authors.initials: B
authors.lastname: LeBlanc
authors.forename: Jutta K
authors.initials: JK
authors.lastname: Preiksaitis
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000382:microbiology; D004279:DNA, Viral / Q000097:blood*; D006801:Humans; D016377:Organ Transplantation / Q000009:adverse effects*; D010750:Phosphoproteins / Q000097:blood*; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012044:Regression Analysis; D012680:Sensitivity and Specificity; D019562:Viral Load; 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