Simultaneous monitoring of CMV and BKV by quantitative PCR in renal transplant recipients

Abstract

Polyomavirus (BKV) and cytomegalovirus (CMV) are associated with renal graft failure. The aim was to establish a quantitative PCR method (Q-PCR) to detect BKV and CMV simultaneously. The conserved sequences of BKV and CMV were amplified and cloned into the plasmids as standards. The sensitivity, specificity and the precision of the assay were evaluated. Q-PCR was used to detect BKV and CMV DNA simultaneously in 480 blood samples of renal transplantation recipients. The sensitivity of the Q-PCR assay to detect BKV or CMV DNA reached 5×103copies/mL. The use of control DNA verified that the assay could specifically detect the target DNA. The precision of the assay to quantify target DNA copies was acceptable (ICV 3.44% for BKV and 2.23% for CMV; differences between batches ICV 4.98% for BKV and 3.76% for CMV). In 480 samples, 130 samples (27.08%) were CMV DNA positive, which was significantly higher than the 64 BKV DNA positive samples (13.33%, p<0.05). BKV or CMV DNA positivity was significantly associated with high concentrations of Tacrolimus (TAC) (p value<0.05). The Q-PCR assay to detect both CMV and BKV DNA simultaneously was developed successfully with high sensitivity, precision, and time-effectiveness for clinical measurement.