Abstract P1-03-03: High concordance of ER, PR, HER2 and Ki67 by central IHC and FISH with mRNA measurements by GeneXpert® breast cancer stratifier assay
Authors: Wu NC , Wong W , Ho KE , Chu VC , Rizo A , Davonport S , Kelly D , Makar R , Jassem J , Duchnowska R , Biernat W , Radecka B , Fujita T , Klein JL , Stonecypher M , Ohta S , Juhl H , Weidler JM , Bates M , Press MF 
Published: March 2017

Abstract
Current methods for the assessment of ER, PR, Her2, and Ki67 using FFPE tissues are hard to standardize and difficult to perform in Low and Middle Income Countries (LMIC). The GeneXpert® breast cancer stratifier assay (RUO) (BC Strat) is a cartridge-based, RT-qPCR assay of ESR1, PGR, ERBB2, and MKi67 mRNAs using FFPE specimens. The assay is fast (&lt;2 hours, including &lt;10 minutes of hands-on time) and easy to perform.
The aims of this study were: 1) to evaluate the concordance of BC Strat using different IHC antibodies and scoring methods in a preliminary dataset (Part I); and 2) to assess concordance between BC Strat and high quality standard methods in an expanded dataset (Part II).
Methods
Part I: IHC Antibody Variability
To assess BC Strat concordance with various IHC antibodies, 155 invasive ductal carcinoma blocks were sourced from 3 sites. Twenty-four adjacent slide sections from each block were prepared and shipped to different labs for BC Strat analysis(Cepheid) or IHC and FISH testing. Table 1 summarizes the IHC antibodies and scoring methods used in each reference lab.
Table 1:IHC antibodies and scoring methods used in reference labs Antibody for IHClabIHC scoringERSP1MPLNAutomated (Aperio)ER6F11Path IncAutomated (Aperio)ER6F11USCManualPRIE2MPLNAutomated (Aperio)PR16Path IncAutomated (Aperio)PRPGR636USCManualHer2*4B5MPLNAutomated (Aperio)Her2*HercepTestUSCManualKi6730-9MPLNAutomated (Aperio)Ki67MIB1Path IncAutomated (Aperio)Ki67MIB1USCManual*HER2 FISH (all with PathVysion kit) was performed at USC
Part II: Concordance Study
522 invasive ductal carcinoma FFPE samples were sourced from 5 sites. All BC Strat analysis was performed at Cepheid and all IHC and FISH was performed in the Press laboratory at USC. Overall percent agreement (OPA), positive percent agreement (PPA), and negative percent agreement (NPA) between BC Strat and IHC were determined.
Results
Part I: IHC Antibody Variability
Table 2 summarizes the OPA for BC Strat analysis and IHC performed with different IHC antibodies and scoring methods. Slightly better concordance for ER and PR was observed between the BC Strat and the IHC methods performed at USC. Discordant IHC results were also observed among the reference labs' standard methodologies.
Table 2: Overall Percent Agreement between IHC and BC Strat Reference LabOPA with BC StratERMPLN92%ERPath Inc96%ERUSC98%PRMPLN84%PRPath Inc83%PRUSC87%Her2*MPLN*93%Her2*USC*91%Ki67MPLN75%Ki67Path Inc67%Ki67USC81%*for IHC 2+(equivocal), FISH HER2/CEP17 ratio was examined
Part II: Concordance Study
Of the 522 samples tested, 499 (96%) yielded valid results for both BC Strat and IHC (IHC and FISH for Her2). OPA between BC Strat and IHC was 98% for ESR1, 91% for PGR, 93% for ERBB2 (IHC and FISH, 97% for Her2 IHC excluding IHC2+), and 81% for MKi67.
Conclusion
BC Strat assay measurements for ESR1, PGR, ERBB2 and MKi67 mRNA expression in FFPE breast tumor tissues are highly concordant with IHC and FISH performed by high quality reference labs. Further investigations using clinical outcomes from independent studies including prospective-retrospective clinical trials are in progress.
Citation Format: Wu NC, Wong W, Ho KE, Chu VC, Rizo A, Davonport S, Kelly D, Makar R, Jassem J, Duchnowska R, Biernat W, Radecka B, Fujita T, Klein JL, Stonecypher M, Ohta S, Juhl H, Weidler JM, Bates M, Press MF. High concordance of ER, PR, HER2 and Ki67 by central IHC and FISH with mRNA measurements by GeneXpert® breast cancer stratifier assay [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-03-03.