Establishment of a competitive PCR for N-myc amplification in neuroblastoma

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N-myc amplification correlates well with the poor prognosis of neuroblastoma. Although the detection of N-myc amplification in neuroblastoma is of great importance, Southern hybridization, being employed solely so far, has some disadvantages such as time consuming, the requirement of larger amount of DNA,and somewhat complicated procedure. In order to overcome the disadvantages, we planned to apply competitive PCR (cPCR) to the measurement of N-myc amplification. Although the standard polymerase chain reaction (PCR) is essentially unsuitable for the quantitative detection of a given template DNA,cPCR is an absolute determination of the dose of a given template DNA.In 1995, we established cPCR system for determining N-myc amplification by employing the competitor plasmid pZH2.In 1996, we examined the availability of cPCR in the copy-number determination using the samples with small amounts. We tested the N-myc copy number in 6 neuroblastoma samples collected by needle biopsy and demonstrated the availability. Taken together, we concluded cPCR established in this study is a rapid, simple, and being applicable to small samples and can be an alternate method of Southern hybridization to determine N-myc copy number.