The aim of this investigation was to evaluate the test performance of the Adenovirus R-gene Assay (bioMerieux) for quantitative measurement of adenovirus in plasma. Adenovirus is a serious complication for immunocompromised patients, leading to high levels of morbidity and mortality. Detection and monitoring of adenovirus infection is therefore important for clinical management of high-risk patients. The Adenovirus R-gene Assay quantitatively detects 52 adenovirus serotypes from plasma, whole blood, CSF, and ocular samples. This method evaluation was performed on whole blood and plasma samples for the intention of testing plasma as the preferred sample type. Total nucleic acids were purified and amplified from samples and controls using the MagNAPure 96/LC480 System (Roche Diagnostics). Four quantitative standards were used to generate a standard curve. The quantitation of samples was extrapolated from the standard curve to provide results between the ranges 500 copies/mL and 1x107 copies/mL. Analytical sensitivity was evaluated using serial dilutions of an adenovirus DNA standard (Vircell). Analytical precision was evaluated with a quantitative adenovirus DNA Control (Acrometrix). The lower limit of detection concurred with the calculated viral load. Ten replicates at 1 x104 copies/mL demonstrated precision of +/- 0.06 log10. Whole blood and plasma samples previously tested using an in-house qualitative adenovirus assay (n=42) were tested using Adenovirus R-gene. For those samples not detected using qualitative in-house testing (n=21), all were concordant with Adenovirus R-gene (<500 copies/mL). For those samples detected (n=21), 20 demonstrated reportable viral loads within the range of 9.44 x 102 to 6.34 x 105 copies/mL. One sample was reported as <500 copies/mL (visible amplification was observed but after the threshold for absolute quantitation). There was insufficient sample to repeat the test. The result may be at the lower limit of detection or the sample may have deteriorated (stored since 2013). Overall the Adenovirus R-gene assay performed well, is suited for routine use, and is able to be simultaneously used with our existing BKV R-gene assay.