Comparison of commercial and in-house Real-time PCR assays for quantification of Epstein-Barr virus (EBV) DNA in plasma

BMC Microbiol. 2007 Mar 28:7:22. doi: 10.1186/1471-2180-7-22.

Abstract

Background: Epstein-Barr virus (EBV) DNA load monitoring is known to be useful for the diagnosis and monitoring of EBV-associated diseases. The aim of this study is to compare the performance of two real-time PCR assays for EBV DNA: a commercial kit as the Q-EBV Real-Time System (Q-EBV PCR, Amplimedical, Turin, Italy) and an in-house assay (EBV RQ-PCR).

Results: The range of linearity and the degree of precision of the two assays were similar. The clinical sensitivity of Q-EBV PCR was higher for reference samples containing less than 1,000 EBV DNA copies/ml. The absolute quantitative results of the two methods were statistically correlated (R2 = 0.7789; p < 0.0001), with the systematic overestimation by EBV RQ-PCR possibly linked to different amplification efficiency in calibration standards.

Conclusion: Both the commercial and the in-house assay may be appropriate for clinical use, but common standards are advisable for comparable absolute values, as these would improve the clinical utility of EBV DNA load measurement.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calibration
  • Cell Line
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Epstein-Barr Virus Infections / diagnosis
  • Epstein-Barr Virus Infections / virology
  • Herpesvirus 4, Human / genetics*
  • Humans
  • Plasma / virology*
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • DNA, Viral