Evaluation of an automated extraction system in combination with Affigene® CMV Trender for CMV DNA quantitative determination: Comparison with nested PCR and pp65 antigen test

CMV viral load quantitation is a powerful tool to assist clinicians in making accurate diagnoses, managing post-transplant CMV disease and monitoring antiviral therapy. The aim of this study was to evaluate the performance of Affigene® CMV Trender for CMV viral load determination used in combination with a non-dedicated nucleic acid extraction system (BioRobot MDx) for high-throughput routine. Linearity, reproducibility and sensitivity were examined. Clinical samples were used to compare results obtained with the Affigene® CMV Trender, with an “in house” nested PCR used for routine diagnosis and with pp65 antigenemia. The results indicated that the test is linear in the range of 1.81–5.18 Log copies/ml and that sensitivity is 77 copies/ml. The concordance of the Affigene® CMV Trender with nested PCR was high, (k = 0.91, IC 95% = 0.82–1.00), whereas a substantial concordance with pp65 antigenemia was observed (k = 0.64, IC 95% = 0.54–0.73). In conclusion, combined use of a non-dedicated automated nucleic acid extraction method with the Affigene® CMV Trender results in an accurate high throughput system, suitable for routine laboratory monitoring of CMV infection.