Competitive polymerase chain reaction for the quantification of N-myc gene copy number in neuroblastoma

An absolute quantification method for the N-myc gene copy number of neuroblastoma specimens was established by applying the competitive polymerase chain reaction (cPCR). The competitor plasmid (pZH2) lacking an MluI site in the exon 2 was constructed to distinguish two product species amplified from genomic DNA and the competitor plasmid. By using this cPCR system, we could obtain qualitative results within 1 day, i.e. amplified or unamplified, and quantitative results by using radiolabelled nucleotides within 4 days. The copy numbers of N-myc in 47 neuroblastoma specimens by cPCR correlated well with those by Southern hybridization (r = 0.85). We conclude that cPCR is a simple and rapid method, requires only a small amount (200 ng) of sample DNA, and is expected to be used for prognostic evaluation in neuroblastomas.