A POLYMERASE CHAIN REACTION-BASED ASSAY FOR THE RAPID DETECTION OF GENE AMPLIFICATION IN HUMAN TUMORS

A differential polymerase chain reaction (PCR) protocol was establishe d for semiquantitative, nonradioactive detection of gene amplification using a DNA sequencer. Oncogene fragments and control DNA sequences w ere simultaneously PCR-amplified using fluorescent-labelled primers. A nalysis of the PCR products allowed a quantitative assessment of gene copy numbers in this coamplification assay, Using this approach, we ex amined a series of 132 brain tumors for amplification of the epidermal growth factor receptor (EGFR) gene. The same set of tumors was also a nalyzed by Southern blotting and hybridization with a radiolabelled EG FR probe. Both methods yielded virtually identical results. This techn ique has a great potential for nonradioactive screening of large tumor panels for oncogene amplification.