Comparison of Real-Time PCR and Immunohistochemistry for HER-2/neu gene amplification assay in breast cancer tissue

AUTHORS

Ahmad Kaviani 1 , Rezvan Esmaeili 2 , Mohsen Foroughizadeh Moghaddam 3 , Nasrin Abdoli 4 , Ali-Akbar Zare 4 , Mohammad Hossein Lashgari 5 , Keivan Majidzadeh 6 , *

1 Associated professor, Department of Surgery, Tehran University of Medical Sciences, Tehran, Iran, Andorra

2 Researcher, Cancer Genetics Research Group, Breast Cancer Research Center (BCRC), ACECR, Tehran, Iran, Andorra

3 Researcher, Novin medical radiation institute, Tehran, Iran, Andorra

4 - Researcher, Cancer Genetics Research Group, Breast Cancer Research Center (BCRC), ACECR, Tehran, Iran, Andorra

5 - Assistant Professor, Department of Surgery, AJA University of Medical Science, Tehran, Iran, Andorra

6 Associated professor, Faculty of Medicine, AJA University of Medical Science, Tehran, Iran and Associated professor, Cancer Genetics Research Group, Breast Cancer Research Center (BCRC), ACECR, Tehran, Iran., Andorra

How to Cite: Kaviani A , Esmaeili R , Foroughizadeh Moghaddam M , Abdoli N , Zare A , et al. Comparison of Real-Time PCR and Immunohistochemistry for HER-2/neu gene amplification assay in breast cancer tissue, Ann Mil Health Sci Res. 2013 ; 11(3):e65138.

ARTICLE INFORMATION

Annals of Military and Health Sciences Research: 11 (3); e65138
Published Online: September 18, 2013
Article Type: Original Article
Received: May 29, 2013
Accepted: August 18, 2013

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Abstract

Background: HER-2/neu is one of the major molecular prognostic markers in breast cancer and due to treatment decision guidelines, the amplification of this gene is very important in breast cancer patients treatment. The most commonly used method for detecting HER-2/neu gene amplification is immunohistochemistry (IHC). Unquantifiable results, lack of reproducibility and long procedure time are among the drawbacks of this method which have persuaded researchers to seek alternative modern molecular based techniques such as Real-Time PCR. The aim of our study was the comparison of HER-2 gene amplification assessed by RealTime PCR and IHC method. Materials and Methods: After DNA extraction, 41 formalin-fixed paraffin-embedded (FFPE) breast carcinoma with known IHC results, had been assessed using Real-Time PCR and gene copy number of HER-2 were measured and compared with the results of IHC. Fluorescent in situ hybridization (FISH) for equivocal IHC and unreliable results were performed. The Fisher's Exact Test and Phi Cramer's V Coefficient were used for statistical analysis. Results: According to the results, 39% of samples tested by Real-Time PCR have HER-2/neu gene amplification and 36% of samples was positive by IHC. The correlation of two methods in positive and negative samples were 85% and 87% respectively. Overrall correlation between two methods was 71%. Also, IHC2+ results have been assessed by FISH technique and all of them were negative, which was in concordance with the results of the Real-Time PCR. Conclusion: About 71% concordance had been observed between two methods. In the case of IHC2+ results, FISH was performed and proved the Real-Time PCR results. Real-Time PCR could precisely measure the amount of tissue HER2 amplification and also were in agreement with IHC results. Since Real-Time PCR has high precision, also is cheaper and more accessible than other methods, we suggest it to be used to assess this indicator.

Keywords

Breast Neoplasms HER-2/neu Copy Number Changes DNA Polymerase Chain Reaction

© 2013, Annals of Military and Health Sciences Research. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

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