Quantitation of CMV by real-time PCR in transfusable RBC units.
BACKGROUND
CMV is one of the most significant pathogens infecting immunocompromised individuals. CMV is transmissible through transfusion of blood components. The goal of this study was to measure CMV levels in RBC units using a sensitive and quantitative DNA amplification assay.


METHODS
An assay to measure CMV load was developed by using real-time PCR to target the major immediate early viral gene. A probe (TaqMan, Applied Biosystems) was used to confirm product specificity and to permit quantitation of CMV in blood samples on a sequence detection system (ABI Prism 7700, Applied Biosystems).


RESULTS
The assay was shown to be accurate, linear, and sensitive to as few as five copies of CMV DNA per PCR. The assay was applied to aliquots of RBC units from 203 healthy donors, 110 of whom were seropositive for CMV. CMV DNA was not detected in any of the 203 RBC samples.


CONCLUSIONS
The findings statistically imply that at least 98.5 percent of RBC units have a CMV load of less than 250 copies per mL. Future clinical studies on larger numbers of units are required to determine the utility of real-time PCR in evaluating the risk of CMV transmission and in confirming the efficacy of WBC reduction.
Greenlee
Deborah J
DJ
University of Texas Health Science Center at San Antonio, USA.
Fan
Hongxin
H
Lawless
Kathleen
K
Harrison
Chantal R
CR
Gulley
Margaret L
ML
D001782:Blood Donors; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000097:blood*; D017707:Erythrocyte Transfusion; D004912:Erythrocytes / Q000821:virology*; D006801:Humans; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D019562:Viral Load
D004279:DNA, Viral