Human cytomegalovirus DNA in plasma and serum specimens of renal transplant recipients is highly fragmented.
Quantitation of cytomegalovirus (CMV) DNA in plasma and serum by PCR is increasingly used to identify patients at risk for developing CMV disease and to monitor the efficacy of antiviral therapy. Although CMV DNA levels are generally interpreted as viral loads, the exact nature of the viral DNA in these specimens is unknown. We studied the state of CMV DNA in plasma and serum specimens obtained from three renal transplant recipients at peak viral DNA levels during primary CMV infection. For this purpose, DNA isolated from these specimens was fractionated by size, and CMV DNA levels in the resulting DNA fractions were measured by quantitative PCR targeted at large (578-bp) and small (134-bp) amplicons. These experiments showed that the molecular sizes of DNA fragments from which CMV DNA is amplified were small (<2,000 bp), indicating that CMV DNA in plasma and serum is highly fragmented. Furthermore, CMV DNA levels were consistently higher when targeted at the smaller amplicon, providing additional evidence for the fragmentation of viral DNA. In conclusion, the first results with three patients have shown that CMV DNA in plasma and serum is highly fragmented and does not necessarily reflect the amount of infectious virus. These observations have potential consequences for understanding CMV pathogenesis and interpreting CMV DNA levels in individual patient management.
Boom
René
R
Section of Clinical Virology, Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Sol
Cees J A
CJ
Schuurman
Tim
T
Van Breda
Alex
A
Weel
Jan F L
JF
Beld
Marcel
M
Ten Berge
Ineke J M
IJ
Wertheim-Van Dillen
Pauline M E
PM
De Jong
Menno D
MD
D000925:Anticoagulants / Q000494:pharmacology; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000821:virology*; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D006801:Humans; D016030:Kidney Transplantation / Q000009:adverse effects*; D008163:Luminescent Measurements; D016133:Polymerase Chain Reaction
D000925:Anticoagulants; D017931:DNA Primers; D004279:DNA, Viral; D004492:Edetic Acid