PATHOLOGY & ONCOLOGY RESEARCHVol. 12 No. 4, 2006

 Article

Detection of HER-2/neu Gene Amplification in Breast Carcinomas Using Quantitative Real-time PCR-A Comparison with Immunohistochemical and FISH Results

Janina KULKA1, Anna-Mária TÕKÉS1, Pál KAPOSI-NOVÁK1, Nóra UDVARHELYI2, Anikó KELLER1, Zsuzsa SCHAFF1

12nd Department of Pathology, Semmelweis University, Budapest, Hungary
2National Institute of Oncology, Budapest, Hungary

 

The aim of our study was to evaluate the value of quantitative real-time-PCR (qPCR) in the determination of HER-2/neu amplification status of human breast carcinomas by comparing qPCR, FISH and immunohistochemistry results from the same samples. A total of 210 breast carcinomas were examined. Ready-to-use CB11 antibody was applied to detect HER-2/neu oncoprotein expression. In 76 out of 210 cases FISH was performed, and 162 cases were investigated with qPCR. Seventy-five tumors were 2+ or 3+ positive with immunohistochemistry, while 135 samples were either completely negative or 1+. In 45 cases results from all three methods were available. Out of these, in twenty negative and sixteen positive cases both FISH and qPCR led to similar results. The mean qPCR amplification ratio in the concordant positive cases was 5.424 while in the qPCR+/FISH- group the mean ratio was 2.765. Out of 121 samples with scores of 0 or 1+ immunohistochemical result, analyzed also with qPCR, 26 showed HER-2/neu gene amplification. In these cases the mean amplification ratio was 2.53. Comparison of FISH and qPCR together with immunohistochemistry shows that qPCR is more sensitive to detect HER-2/neu gene amplification in tumors scored as 2+ with immunohistochemistry, but the diagnostic cut-off ratio should be defined above 2.7 to avoid high number of false positive cases. Amongst the immunohistochemistry score 2+ cases, 10 of 18 showed gene amplification by qPCR while 10 of 26 by FISH. In conclusion, a well calibrated HER-2/neu qPCR assay may serve as useful alternative to FISH in breast cancer patients. Pathology & Oncology Research, Vol 12, Nr 4, 197-204, 2006

Key words: HER-2/neu protein; HER-2/neu DNA amplification; breast carcinoma; immunohistochemistry; FISH; quantitative RT-PCR; LightCycler RT-PCR system


Received: Sep 15, 2006; accepted: Oct 10, 2006
Correspondence: Janina KULKA, 2nd Department of Pathology, Semmelweis University, Üllõi út 93. Budapest H-1091, Hungary; Tel: (36-1) 2157300/3504, Fax: (36-1)2156921; E-mail: kj@korb2.hu

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