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.
Inoue
A
A
Department of Molecular Biology, Toho University School of Medicine, Japan.
Hasan
Z
Z
Hemmi
H
H
Kanda
N
N
Hayashi
Y
Y
Ishizawa
T
T
Tsuchida
Y
Y
Shimatake
H
H
D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics; D018628:Gene Dosage; D016259:Genes, myc; D006801:Humans; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000235:genetics*; D016133:Polymerase Chain Reaction