Quantitative determination of estrogen receptor, progesterone receptor, and HER2 mRNA in formalin-fixed paraffin-embedded tissue--a new option for predictive biomarker assessment in breast cancer.
Author(s): Müller BM, Kronenwett R, Hennig G, Euting H, Weber K, Bohmann K, Weichert W, Altmann G, Roth C, Winzer KJ, Kristiansen G, Petry C, Dietel M, Denkert C
Publication: Diagn Mol Pathol, 2011, Vol. 20, Page 1-10
PubMed ID: 21326033 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of storage, amplicon length, and targeted transcript on real-time RT-PCR success from single sections of FFPE breast tumors. The authors also compared real-time RT-PCR and IHC staining results.
Summary of Findings:
Amplifications of the 67 bp and 151 bp fragments of G6PDH were successful in 100% and 99% of specimens, respectively. In contrast, amplification of the 242 bp fragment was successful in only 1.8% of specimens and amplification of larger fragments was not possible. RNA average yield decreased from 2.9 ug to 1.4 ug per section and the RPL37A CT increased from 22 to 25 over the first 10 years of storage, but no further storage effects were seen. A high degree of concordance was observed between qRT-PCR and IHC for ESR1 (98.4%), PGR (84.4%), and HER2 (89.8%). Different sections from the same tumor showed similar CT values for all three biomarkers, regardless of storage duration.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform RNA Real-time qRT-PCR RNA RT-PCR Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration <1 y
2-5 y
11-13 y
14-16 y
17-21 y
RT-PCR Specific Length of gene fragment 67 bp
151 bp
242 bp
379 bp
453 bp
RT-PCR Specific Targeted nucleic acid Glucose-6-phosphate dehydrogenase
Real-time qRT-PCR Specific Targeted nucleic acid HER2
ESR1
PGR
RPL37a
Immunohistochemistry Specific Targeted peptide/protein HER2
ESR
PGR