Duplex polymerase chain reaction for the simultaneous detection of the human polyomavirus BK and JC DNA

With the aim of further simplifying the detection of human polyomaviruses BK (BKV) and JC (JCV), by reducing the time and the cost of the reactions, a duplex polymerase chain reaction (PCR) was developed for the simultaneous detection and identification of BKV and JCV DNA. A nested PCR was performed. In a first amplification reaction the same set of primers was used to amplify a sequence of either BKV or JCV genomes. Two sets of primers, each specific for BKV or JCV DNA, were used in a second amplification. The products of BKV and JCV DNA amplification were distinguishable on the basis of their different size. The sensitivity and the specificity of the duplex PCR was shown to be exactly comparable to that of the single PCR, not only with the BKV and the JCV plasmid but also for BKV and JCV DNA detection in clinical specimens.