Human cytomegalovirus (CMV) DNA in plasma reflects quantity of CMV DNA present in leukocytes

Abstract

A quantitative DNA amplification assay for human cytomegalovirus (CMV) DNA has been used to evaluate the relationship between quantities of CMV DNA in plasma and those in infected leukocytes (WBC) from human immunodeficiency virus-infected patients. The target sequence for DNA amplification was a region of the immediate-early 1 gene of CMV. The quantitation assay uses an internal control that is coamplified with each patient sample DNA and contains a sequence for detection by colorimetric hybridization with the same bases, but in different order than in the CMV immediate-early 1 region used for hybridization of amplified patient sample DNA. Results showed that patients with CMV disease had more CMV DNA in both WBC and plasma than those without disease. However, in this study, copy numbers of CMV DNA in WBC were higher than those in plasma. The gB and gH variants were the same in plasma and WBC.

Sensitivity of nested PCR toward human cytomegalovirus-DNA in native sera and in extracted DNA of serum samples

“…This may be attributed to the effect of Wizard® DNA purification mini kit in decreasing the effect of inhibitors on detection of HCMV DNA in sera of HCV patients. Similar results were reported by Gerna et al, (1994), Zipeto et al, (1995), Klaus et al, (1997) and Klein et al, (1997).…”

Antibody response to human cytomegalovirus (HCMV) glycoprotein B (gB) in AIDS patients with HCMV end-organ disease

“…Most treated patients did not have antigenemia and viremia, whereas the majority of untreated patients were antigenemic and viremic at the time of sampling. Considering the efficacy of systemic anti-HCMV therapy in reducing blood viral loads [Gerna et al, 1994], and the fact that positive antigenemia and viremia tests reflect the presence of a high systemic viral burden [Gerna et al, 1994;Zipeto et al, 1995], our data suggest strongly the existence of an inverse correlation between the systemic viral loads and the anti-gB and neutralizing antibody levels measurable by our assays. However, validation of this assumption requires a quantitative estimation of viral loads in addition to measuring antibody levels, which unfortunately could not be carried out this study.…”

False-Positive Results of Plasma PCR for Cytomegalovirus DNA due to Delayed Sample Preparation

“…By quantitative PCR techniques, demonstration of high-level CMV leukocyte DNAemia and plasma DNAemia usually precedes the development of clinical features. Although levels of leukocyte DNAemia and plasma DNAemia are usually well correlated, CMV DNA load is considerably higher in PBLs, which can reasonably be explained by the strong cell association of the virus (2,6,18,22). As the performance of quantitative PCR is technically demanding and cost-intensive, which limits its highthroughput use, qualitative CMV PCR assays with reliable significance remain desirable.…”

“…Despite the important diagnostic role of CMV plasma DNAemia, its biological properties are not fully understood. One notion is that it reflects active virus replication (20)(21)(22). On the other hand, the high abundance of CMV leukocyte DNAemia in actively CMV-infected patients has led to the hypothesis that plasma DNAemia may be, at least partly, a result of PBL lysis (6,22).…”

“…One notion is that it reflects active virus replication (20)(21)(22). On the other hand, the high abundance of CMV leukocyte DNAemia in actively CMV-infected patients has led to the hypothesis that plasma DNAemia may be, at least partly, a result of PBL lysis (6,22). If cell turnover was a major mechanism that elicits CMV plasma DNAemia, this would suggest that in patients with CMV leukocyte DNAemia long-term storage of whole blood samples before separation may cause release of viral DNA into the plasma fraction, thus distorting PCR results.…”