Ann Lab Med.  2015 Jan;35(1):99-104. 10.3343/alm.2015.35.1.99.

Comparison of Quantitation of Cytomegalovirus DNA by Real-Time PCR in Whole Blood with the Cytomegalovirus Antigenemia Assay

• Kwon S
• Jung BK
• Ko SY
• Lee CK
• Cho Y

• KMID: 2363155
• DOI: http://doi.org/10.3343/alm.2015.35.1.99

Abstract

BACKGROUND
Quantitation of cytomegalovirus (CMV) DNA using real-time PCR has been utilized for monitoring CMV infection. However, the CMV antigenemia assay is still the 'gold standard' assay. There are only a few studies in Korea that compared the efficacy of use of real-time PCR for quantitation of CMV DNA in whole blood with the antigenemia assay, and most of these studies have been limited to transplant recipients. METHOD: 479 whole blood samples from 79 patients, falling under different disease groups, were tested by real-time CMV DNA PCR using the Q-CMV real-time complete kit (Nanogen Advanced Diagnostic S.r.L., Italy) and CMV antigenemia assay (CINA Kit, ArgeneBiosoft, France), and the results were compared. Repeatedly tested patients were selected and their charts were reviewed for ganciclovir therapy.
RESULTS
The concordance rate of the two assays was 86.4% (Cohen's kappa coefficient value=0.659). Quantitative correlation between the two assays was a moderate (r=0.5504, P<0.0001). Among 20 patients tested repeatedly with the two assays, 13 patients were transplant recipients and treated with ganciclovir. Before treatment, CMV was detected earlier by real-time CMV DNA PCR than the antigenemia assay, with a median difference of 8 days. After treatment, the antigenemia assay achieved negative results earlier than real-time CMV DNA PCR with a median difference of 10.5 days.
CONCLUSIONS
Q-CMV real-time complete kit is a useful tool for early detection of CMV infection in whole blood samples in transplant recipients.

Keyword

Cytomegalovirus; Real-time PCR; Antigenemia assay

MeSH Terms

Antiviral Agents/therapeutic use
Cytomegalovirus/*genetics
Cytomegalovirus Infections/drug therapy/pathology/virology
DNA, Viral/*blood/metabolism
Ganciclovir/therapeutic use
Humans
*Immunoassay
Organ Transplantation
Phosphoproteins/genetics/immunology/*metabolism
*Real-Time Polymerase Chain Reaction
Viral Matrix Proteins/genetics/immunology/*metabolism
Virology/*methods
Antiviral Agents
DNA, Viral
Ganciclovir
Phosphoproteins
Viral Matrix Proteins

Figure

• 

  Fig. 1 Correlation between the cytomegalovirus (CMV) DNA copy number and the number of pp65-antigen-positive cells in 156 real-time CMV DNA PCR-positives. Results from these tests are expressed as follows: pp65 assay as positive cells/200,000 examined white blood cells (WBCs); CMV DNA as log10 copies/mL (Pearson's correlation coefficient; n=156, r=0.5504, P<0.0001). The linear regression (solid line) with 95% confidence intervals (dotted line) is shown.

• 

  Fig. 2 Evaluation of cytomegalovirus (CMV) DNA in whole blood samples for different groups of pp65-antigenemia (Pearson's correlation coefficient; n=156, r=0.3877, P=0.0013). The CMV DNA load was plotted for four pp65-antigenemia groups. For each pp65 group, the median CMV DNA load (log10 copies/mL), the interquartile 50% range, and the range of values are represented. Abbreviation: WBC, white blood cell.

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