Evaluation of a human adenovirus viral load assay using the Altona RealStar® PCR test

Abstract

This study evaluated the performance of the Altona Diagnostics RealStar® Adenovirus Research Use Only (RUO) real-time PCR reagents for HAdV quantitation in plasma samples from immunodeficient patients. The assay was linear from 2.30-9.17 log₁₀ copies/mL (coefficient of determination; R²=0.998) with limits of detection and quantification of 2.19 log₁₀ and 2.30 log₁₀ copies/mL (>95% positivity rate), respectively. Assay precision was highly reproducible with coefficients of variance ranging from 0% to 4.7%. A comparison of 66 matched samples showed good agreement (R²=0.845) between the Altona and the reference laboratory assay, with an average negative bias (-0.24 log₁₀ copies/mL). Genotyping analysis demonstrated that HAdV species B and C accounted for 77% of the positive samples. A significant (≥0.9 log₁₀) difference in quantitation between both tests was found for three HAdV types (HAdV types A12, B14 and F41). In conclusion, the Altona RealStar® test is a reliable and sensitive assay for HAdV DNA quantitation.

Keywords: Adenovirus quantitation; Automation; Real-time PCR test; Viral load.