1773. Development and Evaluation of an Automated Adenovirus Quantitative Assay Using the Luminex ARIES® System

BACKGROUND: The quantification of circulating human adenovirus (HAdV) DNA is the recommended diagnostic method to predict disseminated disease. Most current HAdV quantitative assays are manually performed and lack the flexibility needed to provide rapid answers when required.The purpose of this study was to evaluate the application of MultiCode® HAdV PCR assay for use with the Luminex ARIES(®) system (Luminex Corporation, Austin, TX) as an automated and random access test for the quantitative detection of HAdV DNA in plasma. METHODS: Analytical performance characteristics including assay limit of detection/quantitation (LoD/LoQ), accuracy, and inter-, intra-reproducibility were studied using commercial panels (Exact Diagnostics, Fort Worth, TX). Assay specificity was determined usingHAdV reference strains obtained from the American Type Culture Collection (Manassas, VA) and plasma spiked with related Herpes viruses and pathogens commonly found in the blood. Assay accuracy was verified with analysis of 30 plasma samples spiked with different concentrations of control material that covered the full range of HAdV DNA levels. We also prospectively analyzed 180 plasma samples collected from 102 patients. DNA from all samples were extracted, amplified and detected on a single automated Luminex ARIES® system. RESULTS: The assay has a wide linear range from 2.55 to 9.4 log(10) HAdV DNA copies/mL (coefficient of determination; R(2) = 0.995) with a detection limit of 1.82 log(10) (95% positivity rate), and a limit of quantification of 2.55 log(10) copies/mL. The assay detected HAdV DNA from Adenovirus groups A-F, although slight shifts in Tm peaks were observed. Inter- and intra-assay reproducibility was evaluated using 6 panels of commercial standards, producing variation coefficients of 5% and 2%, respectively. Assay accuracy results reflected a good correlation with a mean difference of 0.10 log(10) copies/mL. The results of the prospective study showed 6/102 (5.8%) patients had HAdV viremia, including 4 (3.9%) patients with a viral load ≥4 log(10)/mL, which might necessitate therapy. CONCLUSION: The HAdV quantitative assay using the Luminex ARIES(®) system provides excellent performance for routine testing with the additional advantage of random access capabilities for urgent testing to identify patients at risk for disseminated disease. DISCLOSURES: All authors: No reported disclosures.