A polymerase chain reaction-based assay for the rapid detection of gene amplification in human tumors

被引：30

作者：

Waha, A ^{[1
]}

Rollbrocker, B ^{[1
]}

Wiestler, OD ^{[1
]}

vonDeimling, A ^{[1
]}

机构：

[1] UNIV KLINIKEN BONN,INST NEUROPATHOL,D-53105 BONN,GERMANY

来源：

DIAGNOSTIC MOLECULAR PATHOLOGY | 1996年 / 5卷 / 02期

关键词：

gene amplification; fluorescence; DNA sequencer; EGFR; differential PCR; semiquantitation;

D O I：

10.1097/00019606-199606000-00010

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 a 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 for oncogene amplification.

引用

页码：147 / 150

页数：4

共 15 条

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