A polymerase chain reaction-based assay for the rapid detection of gene amplification in human tumors.
A differential polymerase chain reaction (PCR) protocol was established for semiquantitative, nonradioactive detection of gene amplification using a DNA sequencer. Oncogene fragments and control DNA sequences were simultaneously PCR-amplified using fluorescent-labelled primers. Analysis of the PCR products allowed quantitative assessment of gene copy numbers in this coamplification assay. Using this approach, we examined a series of 132 brain tumors for amplification of the epidermal growth factor receptor (EGFR) gene. The same set of tumors was also analyzed by Southern blotting and hybridization with a radiolabelled EGFR probe. Both methods yielded virtually identical results. This technique has a great potential for nonradioactive screening of large tumor panels of oncogene amplification.
Waha
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A
Institut für Neuropathologie, Universitätskliniken, Bonn, Germany.
Rollbrocker
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B
Wiestler
O D
OD
von Deimling
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A
D000782:Aneuploidy; D015139:Blotting, Southern; D001932:Brain Neoplasms / Q000235:genetics*; D004247:DNA / Q000032:analysis; D066246:ErbB Receptors / Q000235:genetics / Q000276:immunology; D005784:Gene Amplification / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000737:chemistry; D008969:Molecular Sequence Data; D009857:Oncogenes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D014314:Trisomy
D004247:DNA; D066246:ErbB Receptors