Discordant Results Obtained for Different Methods of HER-2/Neu Testing in Breast Cancer – A Question of Standardization, Automation and Timing

Abstract

Background HER-2/neu positivity is required for the selection of stage IV breast cancer patients for trastuzumab therapy. We compared the results of the recommended immunohistochemistry (IHC) evaluation with the automated ACIS™ IHC system and with fluorescence in situ hybridization (FISH). These HER-2/neu tissue results were correlated with the serum HER-2/neu (sHER-2/neu) levels at the time of metastatic spread. Patients and Methods A total of 61 IHC slides from 30 patients were stained using the HercepTest™. HER-2/neu gene amplification was determined using the Ventana™ FISH assay. sHER-2/neu levels were measured with the Oncogene Science” ELISA kit. The concordance of HER-2/neu results was determined using the concordance index Kappa (κ). Results The best concordance between any IHC and FISH was found for the automated ACIS system (88.5%, κ=0.68, category “good”). The comparison between the manual interpretations and the automated IHC was categorized as “very good” (95.1%, κ=0.85). The median sHER-2/neu level of FISH positive patients was significantly higher (67 ng/mL) than that of FISH negative patients (17 ng/mL, p=0.018). The increase in HER-2/neu positivity comparing tissue to stage IV serum was statistically significant (p=0.001). Conclusions The concordance between conventional IHC and computerized analysis was very good. The number of patients with stage IV breast cancer with an elevated sHER-2/neu level was much higher than HER-2/neu positivity in tissue. This discrepancy is only partially explained by the influence of tumor load. Patients with an elevated sHER-2/neu level and no tissue overexpression should be considered for retesting of tissue or a new biopsy.