ERBB2 gene amplification detected by fluorescent differential polymerase chain reaction in paraffin-embedded breast carcinoma tissues

✍ Scribed by Han-Xiang An; Dieter Niederacher; Matthias W. Beckmann; Uwe-Jochen Göhring; Anton Scharl; Frauke Picard; Claudiavan van Roeyen; Hans-Georg Schnürch; Hans G. Bender

• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: French
• Weight: 991 KB
• Volume: 64
• Category: Article
• ISSN: 0020-7136
• DOI: 10.1002/ijc.2910640502

✦ Synopsis

For quantificative determination of ERBB2 gene amplification in archival human carcinoma specimens we have developed a rapid, non-radioactive approach, which is based on the differential polymerase chain reaction (PCR) and fluorescent DNA technique. Sequences from the ERBB2 gene and from a singlecopy reference gene were amplified simultaneously by PCR, in which one of each primer pair was fluorescently labelled. PCR products were separated by polyacrylamide gel electrophoresis in an automated DNA sequencer and directly quantified after laser activation and emission scanning using appropriate software. This fluorescent differential polymerase chain reaction (fd-PCR) method was used for quantificative determination of ERBB2 gene amplification in I95 formalin-fixed, paraffinembedded breast carcinoma tissues. ERBB2 gene amplification was found in 52 (26%) of these tumors and correlated significantly with tumor size, absence of estrogen receptor (ER) and pS2 expression, but not with absence of progesterone receptor (PR) or presence of epidermal growth factor receptor (EGF-R) expression, lymph-node metastases or grading. In univariate