Comparison of two commercial quantitative PCR assays and correlation with the first WHO International Standard for human CMV

Abstract

Comparability between CMV assays could be facilitated by the first WHO International Standard for human CMV (standard). Standard dilutions were submitted to nucleic acid extraction with Versant kPCR Molecular systems SP or MagNA Pure LC System followed by the kPCR PLX™ CMV DNA (kPCR) or the CMV R-gene™ assay (R-gene), respectively; 139 clinical specimens were tested. Both assays correlated well with the standard (R² > 0.96) and a matrix effect was observed. Quantitative results correlated reasonably between both assays for whole blood (R² = 0.79) and well for other specimen types (R² = 0.93). Quantification differences were within one log₁₀ of the averaged log₁₀ results for 25/27 blood specimens and for 32/33 other specimens. Calibration to the standard did not increase this percentage. In conclusion, results of both assays showed reasonable correlation with each other and good correlation with the standard. Calibration to the standard did not improve comparability of quantitative results.

Keywords: CMV; Molecular testing; PCR; Viral load; WHO standard.