PCR assay with internal control was developed to quantify the her-2 gene dosage in human breast cancer tumor samples. Recombinant plasmid with a fragment of the her-2 gene containing the fragment of T7 phage DNA was designed especially to be used as an internal control in the PCR her-2 gene dosage assay. PCR conditions were optimized for the simultaneous usage of two templates--human genomic DNA and DNA of recombinant plasmid (internal control). The ability of the technology to discriminate a twofold increase of the her-2 gene dosage was demonstrated. Increased level of her-2 gene amplification was observed in 6 of 38 samples investigated. This new simple rapid assay can be an alternative to fluorescence in situ hybridization (FISH) and immunochemistry (ICH) tests for the detection of her-2 amplification in human tumors. This technique may be a useful tool for large randomized, prospective cooperative group trials and may support selection of optimal therapy for breast cancer patients in the future.
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