PERFORMANCE CHARACTERISTICS OF A MULTICODE-RTx® PCR ASSAY FOR THE QUANTITATIVE DETECTION OF CYTOMEGALOVIRUS DNA IN PLASMA AND URINE

Sensitive and reproducible assays for detecting and monitoring viral DNA levels are essential tools for managing patient populations at risk for CMV disease. We report here on the performance characteristics of a quantitative PCR assay for CMV (qCMV-PCR) developed by EraGen Biosciences Inc. (Madison, WI) utilizing their proprietary MultiCode-RTx® technology for detecting PCR products in real-time. One of the advantages of MultiCode-RTx® is that, since tagged-primers rather than probes are used for amplicon detection, amplification efficiency is the sole variable influencing the kinetics of signal generation resulting in improved precision of target quantitation. The qCMV-PCR assay described here enables multiplexed detection of CMV and an extractable internal control target. Quantitation is accomplished via interpolation of results into externally-generated, lot specific, calibration curves. We have established the analytical performance metrics of the qCMV-PCR assay, including the specimen-type specific lower limits of quantitation (LLOQ) and detection (LOD), and delineated the sample storage requirements, reproducibility, specificity and internal control performance of this novel assay. The results of these experiments indicate that this assay is capable of highly reproducible quantitation of CMV DNA in clinical samples over a range of at least 5 logs, and generates results that are concordant with currently available reference methods for assessing CMV viral loads.