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

1. 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

   • Tissue - Breast

   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

   View more