J Korean Med Sci.  2009 Aug;24(4):571-578. 10.3346/jkms.2009.24.4.571.

Comparison of Quantitative Cytomegalovirus Real-time PCR in Whole Blood and pp65 Antigenemia Assay: Clinical Utility of CMV Real-time PCR in Hematopoietic Stem Cell Transplant Recipients

• Choi SM
• Lee DG
• Lim J
• Park SH
• Choi JH
• Yoo JH
• Lee JW
• Kim Y
• Han K
• Min WS
• Shin WS
• Kim CC

• KMID: 1779186
• DOI: http://doi.org/10.3346/jkms.2009.24.4.571

Abstract

Successful preemptive therapy for cytomegalovirus (CMV) infection in transplant patients depends on the availability of sensitive, specific, and timely diagnostic tests for CMV infection. Although the pp65 antigenemia assay has been widely used for this purpose, real-time quantification of CMV DNA has recently been recognized as an alternative diagnostic approach. However, the guidelines for antiviral therapy based on real-time quantitative polymerase chain reaction (RQ-PCR) have yet to be established. From November 2004 to March 2005, a total of 555 whole blood samples from 131 hematopoietic stem cell transplant (HSCT) recipients were prospectively collected. RQ-PCR was conducted using an Artus(R) CMV LC PCR kit (QIAGEN). Both qualitative and quantitative correlations were drawn between the two methods. Exposure to the antiviral agent influenced the results of the two assays. Additionally, the discrepancy was observed at low levels of antigenemia and CMV DNA load. Via ROC curve analysis, the tentative cutoff value for preemptive therapy was determined to be approximately 2x10(4) copies/mL (sensitivity, 80.0%; specificity, 50.0%) in the high risk patients, and approximately 3x10(4) copies/mL (sensitivity, 90.0%; specificity, 70.0%) in the patients at low risk for CMV disease. Further study to validate the optimal cutoff value for the initiation of preemptive therapy is currently underway.

Keyword

Cytomegalovirus; Hematopoietic Stem Cell Transplantation; Antigenemia; Real-time PCR; Cutoff

MeSH Terms

Adolescent
Adult
Child
Child, Preschool
Cytomegalovirus/genetics/*isolation & purification
Cytomegalovirus Infections/*diagnosis/therapy
DNA, Viral/*blood
Female
*Hematopoietic Stem Cell Transplantation
Humans
Infant
Male
Middle Aged
Phosphoproteins/analysis/immunology
Polymerase Chain Reaction/*methods
ROC Curve
Reagent Kits, Diagnostic
Sensitivity and Specificity
Viral Matrix Proteins/analysis/immunology

Figure

• 

  Fig. 1 Correlation between the pp65 antigenemia and CMV RQ-PCR in whole blood sample (Pearson correlation coefficient=0.569). CMV, cytomegalovirus; RQ-PCR, real-time quantitative polymerase chain reaction.

• 

  Fig. 2 Representative serial changes in CMV DNA load by real-time PCR and antigenemia in two patients (A, B) who received antiviral therapy. CMV, cytomegalovirus; GCV, ganciclovir; RQ-PCR, real-time quantitative polymerase chain reaction.

• 

  Fig. 3 Longitudinal trend of antigenemia values and CMV DNA load in two patients (A, B) who developed CMV diseases. The details are provided in the text. CMV, cytomegalovirus; CTL, cytotoxic T lymphocyte; FOS, foscarnet; GCV, ganciclovir; RQ-PCR, real-time quantitative polymerase chain reaction.

• 

  Fig. 4 (A) Receiver-operator characteristic (ROC) curve and table of statistics for some selected thresholds for pp65 antigenemia cutoff value (≥5) for the lower risk patients as a standard. (B) ROC curve and table of statistics for some selected thresholds for pp65 antigenemia cutoff value (≥1) for the high risk patients as a standard. AUC, area under the ROC curve. CMV, cytomegalovirus; CI, confidence interval.

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