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Amplifications of oncogeneerbB-2 and chromosome 20q in breast cancer determined by differentially competitive polymerase chain reaction

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Summary

A new method of measuring gene copy number in small samples of DNA was used to measure amplification of theerbB-2 gene and of chromosome 20q in breast cancers. This method, termed ‘differentially competitive polymerase chain reaction’ (DC-PCR) combines the advantages of two other techniques for measuring amplification by PCR, namely differential PCR and competitive PCR. The DC-PCR methodology was evaluated for sensitivity and specificity by comparing amplification oferbB-2 measured by DC-PCR with that obtained by fluorescencein situ hybridization (FISH) for 42 cases or Southern blotting and/or slot blot analysis for 34 cases. There was over 90 percent concordance with both FISH and Southern blotting and/or slot blot analysis.

DC-PCR was used to further characterize the newly described amplicon at chromosome 20q. By analyzing DNA from 10 breast cancer cell lines at 7 different loci, we identified a potential common region of amplification of approximately 5 centimorgans at chromosome 20q13 bordered by loci D20S52 and RMC20C001-S1. One hundred and seventeen cases of primary breast cancer were evaluated for amplification at these two loci. Amplification at one or more loci, defined as > 1.5 fold higher copy number than that of normal DNA, was found in 25 cases (21%). Sixteen cases were amplified at only one of the two probes (12 cases for RMC20C001-S1 and 4 cases for D20S52), suggesting that the target gene lies between the two markers or that there are two independent target genes within a small chromosome region.

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Author notes

  1. Anne Kallioniemi

    Present address: Tampere University Hospital, P.O. Box 2000, FIN-33521, Tampere, Finland

Authors and Affiliations

  1. Geraldine Brush Cancer Research Institute at California Pacific Medical Center, 2330 Clay Street, 94115, San Francisco, CA, USA

    Guoren Deng, Mei Yu, Ling-Chun Chen, Dan Moore, Wayne Kurisu & Helene S. Smith

  2. Div. of Molecular Cytometry, Dept. of Lab. Medicine, UCSF, 94143, SF, CA, USA

    Anne Kallioniemi & Frederick M. Waldman

  3. Lawrence Berkeley Laboratory, 94720, Berkeley, CA, USA

    Colin Collins

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  1. Guoren Deng
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  2. Mei Yu
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  4. Dan Moore
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Deng, G., Yu, M., Chen, LC. et al. Amplifications of oncogeneerbB-2 and chromosome 20q in breast cancer determined by differentially competitive polymerase chain reaction. Breast Cancer Res Tr 40, 271–281 (1996). https://doi.org/10.1007/BF01806816

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